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Sample records for aerobic blood cultures

  1. Clinical comparison of the isolator and BacT/Alert aerobic blood culture systems.

    PubMed Central

    Hellinger, W C; Cawley, J J; Alvarez, S; Hogan, S F; Harmsen, W S; Ilstrup, D M; Cockerill, F R

    1995-01-01

    The performance characteristics of the Isolator (Wampole Laboratories, Cranbury, N.J.) and the BacT/Alert (Organon Teknika Corporation, Durham, N.C.) aerobic blood culture systems were compared for 6,009 blood culture sets obtained from patients with suspected bloodstream infections. The BacT/Alert aerobic bottle [BTA(O2)] was continuously agitated while it was incubated in 5% CO2 at 36 degrees C; culture plates prepared from the Isolator tube [I(O2)] were incubated in 5% CO2 at 37 degrees C. From 394 blood cultures, 416 clinically significant isolates of bacteria and yeasts were recovered. The overall yields for BTA(O2) and I(O2) were not significantly different (319 versus 336; P = 0.20). I(O2) recovered significantly more staphylococcus (P < 0.05) and yeast isolates (P < 0.01). BTA(O2) recovered significantly more aerobic and facultatively anaerobic gram-negative bacilli (P < 0.05). In blood culture sets which produced growth of the same organisms in both the BTA(O2) and I(O2) systems, the BTA(O2) system detected growth sooner, but more rapid identification was possible with the I(O2) system by virtue of earlier isolation of colonies on solid media. PMID:7665647

  2. Controlled Clinical Comparison of BacT/ALERT Standard Aerobic Medium with BACTEC Standard Aerobic Medium for Culturing Blood

    PubMed Central

    Mirrett, Stanley; Reller, L. Barth; Petti, Cathy A.; Woods, Christopher W.; Vazirani, Bindu; Sivadas, Rekha; Weinstein, Melvin P.

    2003-01-01

    Standard aerobic media are widely used for culturing blood with the BacT/ALERT (BioMérieux, Inc., Durham, N.C.) (BM) and BACTEC 9240 (BD Diagnostic Systems, Sparks, Md.) (BD) automated continuously monitoring instrument systems. Although similarly composed of soybean-casein digest broths, the formulations of the standard aerobic media available for these instruments differ from each other in supplements and in sodium polyanetholesulfonate concentration. Therefore, we compared the standard aerobic media available for these systems at two university hospitals. Blood samples from adult patients with suspected bloodstream infection were inoculated at the bedside into nonvented BM and BD standard aerobic blood culture bottles and incubated in their respective instruments. The laboratories received 6,743 pairs of bottles that were each filled with 8 to 12 ml of blood. A total of 523 isolates representing true infections were recovered from 257 patients; of these isolates, 348 were recovered from both the BD and the BM bottles, 108 were recovered from the BM bottles only, and 67 were recovered from the BD bottles only (P < 0.005). More staphylococci (P < 0.05), especially coagulase-negative staphylococci (P < 0.05), and yeasts (P < 0.01) were recovered from BM bottles than from BD bottles. Of 291 unimicrobial episodes of bloodstream infection, 220 were detected with both bottles, 41 were detected with the BM bottles only, and 30 were detected with the BD bottles only (difference not significant). Among 335 cultures that were positive in both bottles within the first 72 h of incubation, the median times to detection were 14 h for BM bottles and 13 h for BD bottles. Rates for false-positive results were 0.5% for BM bottles and 0.1% for BD bottles. One BM bottle and seven BD bottles yielded false-negative results. We conclude that the BM medium provides improved recovery of microorganisms, especially staphylococci and yeasts, compared with that provided by the BD medium

  3. Correlation of growth of aerobic blood cultures in hypertonic broth with antibiotic therapy.

    PubMed Central

    Eng, J; Maeland, A

    1982-01-01

    The aim of this study was to elucidate the mechanisms by which sucrose improves growth in a hypertonic medium for isolating aerobes from blood. Clinical blood cultures were made routinely in duplicate in plain broth consisting of brain heart infusion broth with sodium polyanetholesulfonate, gelatin, and penicillinase and the same broth with 20% sucrose added. The growth patterns of Staphylococcus aureus and Enterobacteriaceae from plain and from hypertonic broth were correlated with the presence or absence of antimicrobial therapy in patients when the blood cultures were collected. In S. aureus bacteremias, 58.7% of the positive cultures collected during treatment of patients with beta-lactam antibiotics showed earlier growth or growth only in hypertonic broth, compared with 16.7% of the cultures taken during treatment with other antimicrobial agents (P less than 0.05) and 17.6% of the cultures made in antibiotic-free intervals (P less than 0.01). In the group of cultures yielding growth of Enterobacteriaceae, growth occurred earlier or solely in hypertonic broth in 28.9% of the cultures taken during treatment with beta-lactam antibiotics, compared with 15.7% of the cultures taken during treatment with other antimicrobial agents and 21.6% of the cultures collected in antibiotic-free intervals (differences not statistically significant). It is concluded that treatment with beta-lactam antibiotics is an important reason for the improved growth of S. aureus from hypertonic broth, but other factors are also involved. PMID:7153339

  4. Controlled Comparison of BacT/ALERT FAN Aerobic Medium and BACTEC Fungal Blood Culture Medium for Detection of Fungemia

    PubMed Central

    McDonald, L. Clifford; Weinstein, Melvin P.; Fune, Jose; Mirrett, Stanley; Reimer, Larry G.; Reller, L. Barth

    2001-01-01

    Yeasts are an increasingly common cause of nosocomial bloodstream infections. Methods for their detection are many; controlled comparisons are few. The vented FAN aerobic blood culture medium has been shown to be superior to the standard BacT/ALERT aerobic medium for the detection of fungemia as well as bacteremia. The BACTEC selective fungal medium (FM) (BD Biosciences, Sparks, Md.) allowed detection of more episodes of fungemia than did a resin-containing medium with equal volumes of blood cultured. Therefore, we compared vented FAN to FM for the ability to recover fungi from the blood of patients who were at increased risk of having fungemia. From 5,109 cultures processed for which both FAN and FM bottles were adequately filled, fungi were recovered from 87 cultures. Of these, 47 were detected with both bottles, 12 were detected with FAN only, and 28 were detected with FM only (P < 0.05). FAN was the first bottle positive for 36 of the 47 cultures for which both bottles yielded the same fungus, whereas the FM bottle was the first bottle positive for 11 cultures (P < 0.001). A total of 54 episodes of fungemia were identified, with 40 detected by both media, 4 detected only by FAN, and 10 detected only by FM (P value, not significant). We conclude that the vented FAN aerobic bottle is comparable to the FM bottle for detection of episodes of yeast infection but has the added benefit of detecting bacteria. PMID:11158118

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

    PubMed

    McGowan, J E; Metchock, B G

    1992-04-01

    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

  6. Evaluation of a Plastic Nonvented Aerobic Blood Culture Bottle for Use with the BacT/ALERT Microbial Detection System

    PubMed Central

    Snyder, J. W.; Munier, G. K.; Bostic, G. D.; Bozigar, P. S.; Hanna, R.

    2002-01-01

    The current BacT/ALERT SA (BTA SA) aerobic blood culture bottle is made from glass, does not require venting, and contains a liquid emulsion sensor (LES). Its performance has been shown to be equivalent to that of the vented standard aerobic culture bottle. A further-improved version of the BTA SA bottle, designated the BacT/ALERT plastic SA (BTA PSA) culture bottle, is made from clear plastic to prevent breakage, does not require venting, and contains a modified LES (LES 2) to reduce the possibility of false positives. The BTA PSA provides a practical alternative to the current glass version of this bottle. The plastic bottle is also comparable to the current glass bottle in transparency and growth performance and additionally minimizes the exposure to infectious agents due to glass bottle breakage. PMID:12454188

  7. Controlled evaluation of BacT/Alert standard aerobic and FAN aerobic blood culture bottles for detection of bacteremia and fungemia.

    PubMed Central

    Weinstein, M P; Mirrett, S; Reimer, L G; Wilson, M L; Smith-Elekes, S; Chuard, C R; Joho, K L; Reller, L B

    1995-01-01

    A new medium, FAN, designed to enhance the recovery of microorganisms, has been developed for the BacT/Alert blood culture system (Organon Teknika Corp., Durham, N.C.). We compared the yield and speed of detection of microorganisms in 6,847 adequately filled paired aerobic standard and FAN bottles at four university hospitals. Of 499 clinically significant microorganisms isolated from one or both bottles, significantly more Staphylococcus aureus isolates (P < 0.001), coagulase-negative staphylococci (P < 0.001), yeasts (P < 0.01), and all microorganisms combined (P < 0.001) were recovered from the FAN bottles. Overall, the speeds of detection of positive cultures did not differ between the two medium formulations; mean times to detection in the standard and FAN bottles were 16.1 and 16.0 h, respectively. When a subset of patients on antimicrobial therapy was evaluated, significantly enhanced yield from the FAN bottle was evident for staphylococci. Overall, the FAN bottle outperformed the standard aerobic BacT/Alert bottle. PMID:7790471

  8. Cross-Canada survey of resistance of 2747 aerobic blood culture isolates to piperacillin/tazobactam and other antibiotics

    PubMed Central

    Forward, Kevin R; Franks, Patricia A; Low, Donald E; Rennie, Robert; Simor, Andrew E

    1998-01-01

    OBJECTIVE: To compare the activity of piperacillin/tazobactam with that of other broad parenteral antibiotics against aerobic and facultative anaerobic blood culture isolates in a Canada-wide survey. DESIGN: Fifty-eight laboratories in nine provinces each contributed up to 50 consecutive clinically significant aerobic and facultative anaerobic isolates for susceptibility testing. SETTING: Participating hospitals included both tertiary care and community hospitals. MATERIALS AND METHODS: Testing was performed in five regional centres by using the same microbroth dilution method, and results were interpreted according to National Commitee for Clinical Laboratory Standards M7-A3 and M100-S5 guidelines. RESULTS: Piperacillin/tazobactam and imipenem were both active against more than 99% of the 1616 strains of Enterobacteriaceae species tested. The minimum inhibitory concentration of 90% of isolates (MIC90) of all Enterobacteriaceae species was 2 mg/L for piperacillin/tazobactam compared with 64 mg/L for piperacillin alone. Seventeen per cent of strains of Enterobacteriaceae species were susceptible to piperacillin/tazobactam but resistant to piperacillin. Piperacillin/tazobactam was highly active against Pseudomonas aeruginosa, inhibiting 99.1% of strains. MIC90 was 8 mg/L. Nine per cent of P aeruginosa strains were not susceptible to imipenem. Most of these strains had a MIC of 8 mg/L, which falls in the intermediate category. Ninety-seven per cent of P aeruginosa were susceptible to ciprofloxacin and 97.3% to tobramycin. Ninety-six per cent of strains of Actinobacter species were susceptible to piperacillin/tazobactam, whereas only 76% of strains were susceptible to piperacillin alone. Overall, piperacillin/tazobactam was the most active agent tested; 98% of all strains were susceptible, followed closely by imipenem, to which 97.8% of strains were susceptible. CONCLUSIONS: Aerobic blood culture isolates from Canadian centres continue to be highly susceptible to a

  9. Comparison of the BacT/Alert FAN aerobic and the Difco ESP 80A aerobic bottles for pediatric blood cultures.

    PubMed Central

    Welby-Sellenriek, P L; Keller, D S; Ferrett, R J; Storch, G A

    1997-01-01

    We compared the BacT/Alert system using the aerobic FAN bottle with the ESP system using the 80A aerobic bottle for the detection of pediatric bloodstream pathogens at a children's hospital. From 6,636 blood culture sets complying with the inclusion criteria, 308 pathogens were detected, including 177 that were detected by both systems, 69 that were detected by BacT/Alert FAN only, and 62 that were detected by ESP 80A only (P = 0.6; not significant). BacT/Alert FAN detected more isolates of Staphylococcus aureus (47 versus 34; P = 0.02), while ESP 80A detected more episodes of streptococcal and enterococcal infection. BacT/Alert FAN detected more pathogens from patients receiving antibiotic therapy (107 versus 93; P = 0.04). Of 248 separate episodes of bacteremia or fungemia, 146 were detected by both systems, 56 were detected by ESP 80A only, and 46 were detected by BacT/Alert FAN only (P = 0.37; not significant). The median times to detection were 13.6 h for ESP 80A and 15.7 h for BacT/Alert FAN (P < 0.001). Both systems were considered easy to operate and were free from significant mechanical difficulties. False-positive or false-negative signals were rare or nonexistent with both systems. We conclude that both systems rapidly detect a broad range of pediatric bloodstream pathogens. BacT/Alert FAN provides better detection of Staphylococcus aureus, especially from patients receiving antibiotics. ESP 80A provides better detection of streptococci and enterococci. PMID:9114401

  10. Blood culture

    MedlinePlus

    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 ...

  11. Controlled Clinical Laboratory Comparison of Two Supplemented Aerobic and Anaerobic Media Used in Automated Blood Culture Systems To Detect Bloodstream Infections

    PubMed Central

    Ziegler, R.; Johnscher, I.; Martus, P.; Lenhardt, D.; Just, H.-M.

    1998-01-01

    A 20-ml blood sample was collected from adult patients with suspected bloodstream infections and distributed equally into the four volume-controlled bottles of a blood culture set consisting of aerobic and anaerobic BACTEC Plus/F bottles and aerobic and anaerobic BacT/Alert FAN bottles. All bottles were incubated in their respective instruments for a standard 5-day protocol or until the instruments signalled positivity. Samples in all bottles with negative results by these instruments were terminally subcultured. A total of 8,390 blood culture sets were obtained during the study period, of which 4,402 (52.5%) met the study criteria. Of these, 946 (21.5%) were positive either by instrument signal or by additional terminal subculture of all negative bottles and yielded growth of microorganisms. Five hundred eighty-nine (13.4%) blood culture sets were considered to have recovered 663 clinically significant organisms. When both the BACTEC and the BacT/Alert systems were used, 465 positive sets were detected; BACTEC alone detected 52 positive sets and BacT/Alert alone detected 72 (P = 0.09). No differences were found between the two systems in microbial recovery rate from blood cultures obtained from patients on antibiotic therapy. Significantly more members of the family Enterobacteriaceae (P < 0.01) were detected from patients without antimicrobial therapy by BacT/Alert than by BACTEC. The false-negative rates were 0.20% for BACTEC and 0.32% for BacT/Alert. A significantly higher false-positive rate was found for BACTEC (P < 0.0001). Both systems were comparable for the time to detection of microorganisms. However, gram-positive bacteria were detected faster by BACTEC and Enterobacteriaceae were detected faster on average by BacT/Alert. We concluded that both systems are comparable in their abilities to recover aerobic and anaerobic organisms from blood cultures and a terminal subculture might not be necessary for either of the two systems. The increased positivity

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

    PubMed Central

    McDonald, L C; Fune, J; Gaido, L B; Weinstein, M P; Reimer, L G; Flynn, T M; Wilson, M L; Mirrett, S; Reller, L B

    1996-01-01

    Two recent multicenter blood culture studies found that BacT/Alert FAN (FAN) bottles (Organon Teknika, Durham, N.C.) had increased yields in detecting bacteremia and fungemia compared with standard BacT/Alert (STD) bottles. Because the clinical importance of this increase in microbial recovery is unknown, we performed a retrospective analysis to determine the frequency with which FAN bottles were the sole means of detecting an episode of bacteremia. There were 1,047 positive blood cultures in which both study bottles were adequately filled and the organism isolated was judged to be the cause of sepsis: 240 (23%) were positive only in FAN bottles and 73 (7%) were positive only in STD bottles. Of a total of 664 episodes of bacteremia, 126 (19%) were identified only by FAN bottles and 43 (7%) were identified only by STD bottles (P < 0.0001). Episodes detected only by FAN bottles more often were recurrent events (23 of 126, or 18%) than episodes detected only by STD bottles (2 of 43, or 5%) (P < 0.05) and more commonly occurred in patients receiving theoretically effective antibiotic therapy (33 of 126 [26%] versus 4 of 43 [9%]) (P < 0.05). The medical records for patients with 127 of these episodes (92 FAN bottles only; 35 STD bottles only) were available for review. More than half of both FAN bottle-only (60 of 92, or 65%) and STD bottle-only (20 of 35, or 57%) episodes were judged to be clinically important. We conclude that FAN bottles improve the detection of bacteremia and that the majority of the additional episodes detected are clinically important. The benefits of the greater yield in specific patient populations must be balanced against the higher costs of FAN bottles. PMID:8862581

  13. Blood Culture Test

    MedlinePlus

    ... used to detect the presence of bacteria or fungi in the blood, to identify the type present, ... blood cultures to detect and identify bacteria and fungi. Other related tests that may be performed include: ...

  14. Aerobic fitness is associated with greater hippocampal cerebral blood flow in children.

    PubMed

    Chaddock-Heyman, Laura; Erickson, Kirk I; Chappell, Michael A; Johnson, Curtis L; Kienzler, Caitlin; Knecht, Anya; Drollette, Eric S; Raine, Lauren B; Scudder, Mark R; Kao, Shih-Chun; Hillman, Charles H; Kramer, Arthur F

    2016-08-01

    The present study is the first to investigate whether cerebral blood flow in the hippocampus relates to aerobic fitness in children. In particular, we used arterial spin labeling (ASL) perfusion MRI to provide a quantitative measure of blood flow in the hippocampus in 73 7- to 9-year-old preadolescent children. Indeed, aerobic fitness was found to relate to greater perfusion in the hippocampus, independent of age, sex, and hippocampal volume. Such results suggest improved microcirculation and cerebral vasculature in preadolescent children with higher levels of aerobic fitness. Further, aerobic fitness may influence how the brain regulates its metabolic demands via blood flow in a region of the brain important for learning and memory. To add specificity to the relationship of fitness to the hippocampus, we demonstrate no significant association between aerobic fitness and cerebral blood flow in the brainstem. Our results reinforce the importance of aerobic fitness during a critical period of child development. PMID:27419884

  15. Relevance of Routine Use of the Anaerobic Blood Culture Bottle▿

    PubMed Central

    Grohs, Patrick; Mainardi, Jean-Luc; Podglajen, Isabelle; Hanras, Xavier; Eckert, C.; Buu-Hoï, A.; Varon, E.; Gutmann, Laurent

    2007-01-01

    Using the BacT/Alert automated system, we conducted a 1-year retrospective study on blood cultures, focusing on the relevance of routine use of the anaerobic bottle. The rate of patients with positive blood cultures was 19.7%. Among these, 13.5% had a positive anaerobic bottle in the absence of any aerobic bottle, and 2/3 of these grew with nonobligate anaerobes. These patients were hospitalized in 20 out of 26 wards of the hospital group. For 65.4% of the monomicrobial-positive blood cultures growing Enterobacteriaceae, the anaerobic bottle detected growth earlier than the corresponding aerobic bottle. These data suggest that, in our institution, the use of an anaerobic bottle is still relevant. PMID:17581942

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

    PubMed Central

    Rohner, P; Pepey, B; Auckenthaler, R

    1997-01-01

    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 < 0.0001), other gram-negative bacteria (P = 0.004), and yeasts (P < 0.0001) were isolated from Plus aerobic/F medium, but more streptococci (P < 0.0001), E. coli (P = 0.02) strains and anaerobes (P < 0.0001) were detected with Lytic anaerobic/F medium. Lytic anaerobic/F vials were significantly (P < 0.0001) more often positive at least 6 h before Plus aerobic/F vials (n = 112 versus 52, respectively). Significantly more (P < 0.0001) Plus 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

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

    PubMed

    Rohner, P; Pepey, B; Auckenthaler, R

    1997-10-01

    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 < 0.0001), other gram-negative bacteria (P = 0.004), and yeasts (P < 0.0001) were isolated from Plus aerobic/F medium, but more streptococci (P < 0.0001), E. coli (P = 0.02) strains and anaerobes (P < 0.0001) were detected with Lytic anaerobic/F medium. Lytic anaerobic/F vials were significantly (P < 0.0001) more often positive at least 6 h before Plus aerobic/F vials (n = 112 versus 52, respectively). Significantly more (P < 0.0001) Plus 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

  18. Novel method for detecting micro-organisms in blood cultures.

    PubMed Central

    Sawhney, D; Hinder, S; Swaine, D; Bridson, E Y

    1986-01-01

    A method for detecting the growth of micro-organisms in blood culture by a visual signal is described. The system utilises a single blood culture medium that has been specifically formulated to support growth of aerobic, anaerobic, and microaerophilic micro-organisms. The system is based on the principle that when micro-organisms grow in the medium in a sealed bottle their metabolic products create positive pressure. This positive pressure displaces the infected blood and broth into an upper chamber, which acts as a visual signal of microbial activity. All the test micro-organisms, when inoculated at less than 20 colony forming units into simulated human blood cultures, gave a positive signal. PMID:3098802

  19. Controlled clinical comparison of three commercial blood culture systems.

    PubMed

    Frank, U; Malkotsis, D; Mlangeni, D; Daschner, F D

    1999-04-01

    In a controlled clinical comparison, three commercial blood culture systems--the standard aerobic BacT/Alert bottle (STD), the aerobic BacT/Alert FAN bottle (FAN) and the Isolator system (ISO; Wampole Laboratories, USA) were compared for their ability to detect aerobic and facultatively anaerobic microorganisms. A total of 945 BacT/Alert (STD and FAN) blood culture sets were compared. Of these, 110 blood culture sets (11.6%) yielded growth of 116 clinically significant bacterial and fungal isolates. Microorganisms were recovered from 10.7% (101/945) of the FAN bottles compared to 8.9% (84/945) of the STD bottles. Of the significant isolates, 78 (67.2%) were recovered by both bottles, 29 (25%) by the FAN bottle only and nine (7.8%) by the STD bottle only (P<0.01). Along with 56.1% (530/945) of BacT/Alert blood culture sets, a concomitant ISO tube was obtained. Of the triple (STD + FAN + ISO) blood culture sets, 54 (10.2%) yielded growth of 59 clinically relevant isolates. Microorganisms were detected in 9.1% (48/530) of the FAN bottles, 8.3% (44/530) of the STD bottles and 4% (21/530) of the ISO tubes (P<0.001). Overall, the BacT/Alert system detected more clinically significant microorganisms than the ISO tube; the STD and the FAN bottle each recovered significantly more staphylococci (P<0.01 and P<0.001, respectively) and gram-negative rods (P<0.01, both). In conclusion, the BacT/Alert FAN bottle performed better than the BacT/Alert STD bottle; both BacT/Alert bottles, however, were superior to the ISO tube in terms of recovery of clinically significant microorganisms, including gram-positive and gram-negative bacteria. PMID:10385012

  20. Effects of aerobic exercise on blood pressure and lipids in overweight hypertensive postmenopausal women

    PubMed Central

    Ammar, Tarek

    2015-01-01

    Menopause may increase risk of hypertension and abnormal lipid profile. The aim of the study was to examine the effects of morning and afternoon aerobic exercises on hypertension and lipids in overweight hypertensive postmenopausal women. Forty five women aged from 49 to 60 years were randomly assigned into three groups. Group (A) 15 patients received medicine, (B) 15 patients performed morning aerobic exercises and received medicine, and group (C) 15 patients performed afternoon aerobic exercises and received medicine. Blood pressure measurement and lipid profile tests were performed before and after the study. The results showed that there was a statistical significant difference among all groups in systolic and diastolic blood pressure, favoring group C. Also there was a statistical significant difference among all groups in lipid levels, favoring group C. Therefore, it can be concluded that morning aerobic exercises were more effective in reducing the blood pressure and lipids than afternoon exercises in overweight hypertensive postmenopausal women. PMID:26171380

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

    SciTech Connect

    Hopfer, R.L.; Orengo, A.; Chesnut, S.; Wenglar, M.

    1980-09-01

    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.

  2. Effect of Aerobic Exercise Training on Blood Pressure in Indians: Systematic Review

    PubMed Central

    Punia, Vandana

    2016-01-01

    Introduction. High blood pressure (BP) is one of the most important modifiable risk factors for cardiovascular diseases, which accounts for one in every eight deaths worldwide. It has been predicted that, by 2020, there would be 111% increase in cardiovascular deaths in India. Aerobic exercise in the form of brisk walking, jogging, running, and cycling would result in reduction in BP. Many meta-analytical studies from western world confirm this. However, there is no such review from Indian subcontinent. Objective. Our objective is to systematically review and report the articles from India in aerobic exercise on blood pressure. Methodology. Study was done in March 2016 in Google Scholar using search terms “Aerobic exercise” AND “Training” AND “Blood pressure” AND “India.” This search produced 3210 titles. Results. 24 articles were identified for this review based on inclusion and exclusion criteria. Total of 1107 subjects participated with median of 25 subjects. Studies vary in duration from +3 weeks to 12 months with each session lasting 15–60 minutes and frequency varies from 3 to 8 times/week. The results suggest that there was mean reduction of −05.00 mmHg in SBP and −03.09 mmHg in DBP after aerobic training. Conclusion. Aerobic training reduces the blood pressure in Indians. PMID:27493989

  3. Effect of Aerobic Exercise Training on Blood Pressure in Indians: Systematic Review.

    PubMed

    Punia, Sonu; Kulandaivelan, Sivachidambaram; Singh, Varun; Punia, Vandana

    2016-01-01

    Introduction. High blood pressure (BP) is one of the most important modifiable risk factors for cardiovascular diseases, which accounts for one in every eight deaths worldwide. It has been predicted that, by 2020, there would be 111% increase in cardiovascular deaths in India. Aerobic exercise in the form of brisk walking, jogging, running, and cycling would result in reduction in BP. Many meta-analytical studies from western world confirm this. However, there is no such review from Indian subcontinent. Objective. Our objective is to systematically review and report the articles from India in aerobic exercise on blood pressure. Methodology. Study was done in March 2016 in Google Scholar using search terms "Aerobic exercise" AND "Training" AND "Blood pressure" AND "India." This search produced 3210 titles. Results. 24 articles were identified for this review based on inclusion and exclusion criteria. Total of 1107 subjects participated with median of 25 subjects. Studies vary in duration from +3 weeks to 12 months with each session lasting 15-60 minutes and frequency varies from 3 to 8 times/week. The results suggest that there was mean reduction of -05.00 mmHg in SBP and -03.09 mmHg in DBP after aerobic training. Conclusion. Aerobic training reduces the blood pressure in Indians. PMID:27493989

  4. Clinical comparison of BACTEC 9240 plus aerobic/F resin bottles and the isolator aerobic culture system for detection of bloodstream infections.

    PubMed Central

    Cockerill, F R; Reed, G S; Hughes, J G; Torgerson, C A; Vetter, E A; Harmsen, W S; Dale, J C; Roberts, G D; Ilstrup, D M; Henry, N K

    1997-01-01

    The Plus Aerobic/F resin bottle of the BACTEC 9240 automated blood culture system (Becton Dickinson Diagnostic Instrument Systems, Sparks, Md.) was compared with aerobic culture of the Isolator system (Wampole Laboratories, Cranbury, N.J.) for the detection of bloodstream microorganisms from 6,145 blood cultures collected from adult patients with suspected septicemia. The BACTEC resin bottles were incubated for 7 days, and the sediment from the Isolator tube was inoculated to sheep blood and chocolate agars which were incubated for 72 h and to inhibitory mold, brain heart infusion, and Sabouraud agars which were incubated for 21 days. A total of 622 microorganisms were recovered from 583 blood cultures. The BACTEC resin bottle recovered statistically significantly more pathogens overall than the Isolator system (P = 0.0006). When individual pathogens isolated from either system for a 7-day study period were assessed, it was determined that the BACTEC resin bottle detected statistically significantly more isolates of Staphylococcus aureus (P = 0.0113) and coagulase-negative Staphylococcus spp. (P = 0.0029) than the Isolator system. The BACTEC resin bottle also detected statistically significantly more bloodstream infections (septic episodes) caused by coagulase-negative Staphylococcus spp. (P = 0.0146). The Isolator system recovered statistically significantly more contaminants overall (P < 0.0001), and among this group of microorganisms, recovered statistically significantly more Bacillus spp. (P < 0.0001), coagulase-negative Staphylococcus spp. (P < 0.0001), and viridans group Streptococcus spp. (P = 0.0156). The Isolator system detected statistically significantly more isolates of Histoplasma capsulatum (P = 0.004), but all of these isolates were detected at > or = 7 days of incubation of fungal plates, i.e., after the system to system comparison study period (7 days). In blood culture sets which produced growth of the same pathogen in both systems, there was a

  5. Clinical comparison of BACTEC 9240 plus aerobic/F resin bottles and the isolator aerobic culture system for detection of bloodstream infections.

    PubMed

    Cockerill, F R; Reed, G S; Hughes, J G; Torgerson, C A; Vetter, E A; Harmsen, W S; Dale, J C; Roberts, G D; Ilstrup, D M; Henry, N K

    1997-06-01

    The Plus Aerobic/F resin bottle of the BACTEC 9240 automated blood culture system (Becton Dickinson Diagnostic Instrument Systems, Sparks, Md.) was compared with aerobic culture of the Isolator system (Wampole Laboratories, Cranbury, N.J.) for the detection of bloodstream microorganisms from 6,145 blood cultures collected from adult patients with suspected septicemia. The BACTEC resin bottles were incubated for 7 days, and the sediment from the Isolator tube was inoculated to sheep blood and chocolate agars which were incubated for 72 h and to inhibitory mold, brain heart infusion, and Sabouraud agars which were incubated for 21 days. A total of 622 microorganisms were recovered from 583 blood cultures. The BACTEC resin bottle recovered statistically significantly more pathogens overall than the Isolator system (P = 0.0006). When individual pathogens isolated from either system for a 7-day study period were assessed, it was determined that the BACTEC resin bottle detected statistically significantly more isolates of Staphylococcus aureus (P = 0.0113) and coagulase-negative Staphylococcus spp. (P = 0.0029) than the Isolator system. The BACTEC resin bottle also detected statistically significantly more bloodstream infections (septic episodes) caused by coagulase-negative Staphylococcus spp. (P = 0.0146). The Isolator system recovered statistically significantly more contaminants overall (P < 0.0001), and among this group of microorganisms, recovered statistically significantly more Bacillus spp. (P < 0.0001), coagulase-negative Staphylococcus spp. (P < 0.0001), and viridans group Streptococcus spp. (P = 0.0156). The Isolator system detected statistically significantly more isolates of Histoplasma capsulatum (P = 0.004), but all of these isolates were detected at > or = 7 days of incubation of fungal plates, i.e., after the system to system comparison study period (7 days). In blood culture sets which produced growth of the same pathogen in both systems, there was a

  6. Clinical value of anaerobic blood culture: a retrospective analysis of positive patient episodes

    PubMed Central

    James, P.; Al-Shafi, K.

    2000-01-01

    Aim—To investigate the clinical value of anaerobic blood culture. Methods—Blood culture bottles (n = 25 185) submitted for culture over a two year period were reviewed. Results—The bottles yielded 1992 positive patient episodes, a positive rate of 14.4/1000 hospital admissions. Significantly more isolations were obtained from aerobic than from anaerobic bottles. Twelve of the 38 anaerobic episodes were detected in aerobic bottles. Clinical management was influenced in one of 24 patients whose cultures yielded anaerobes from anaerobic bottles only. For a further six patients it was unlikely that the result had any effect on clinical management. Conclusions—If aerobic bottles were substituted for the anaerobic bottles, detection of positive patient episodes would increase by at least 6%. A higher yield would be achieved by using two aerobic bottles for routine culture and using anaerobic bottles only for patients where anaerobic culture may influence clinical management. Key Words: blood culture • anaerobes • BacT/Alert PMID:10823145

  7. Vertebrate blood cell volume increases with temperature: implications for aerobic activity

    PubMed Central

    Zenil-Ferguson, Rosana

    2014-01-01

    Aerobic activity levels increase with body temperature across vertebrates. Differences in these levels, from highly active to sedentary, are reflected in their ecology and behavior. Yet, the changes in the cardiovascular system that allow for greater oxygen supply at higher temperatures, and thus greater aerobic activity, remain unclear. Here we show that the total volume of red blood cells in the body increases exponentially with temperature across vertebrates, after controlling for effects of body size and taxonomy. These changes are accompanied by increases in relative heart mass, an indicator of aerobic activity. The results point to one way vertebrates may increase oxygen supply to meet the demands of greater activity at higher temperatures. PMID:24765580

  8. Ralstonia pickettii traced in blood culture bottles.

    PubMed

    Boutros, Névine; Gonullu, Nevriye; Casetta, Anne; Guibert, Michèle; Ingrand, Didier; Lebrun, Léa

    2002-07-01

    Over a 9-month period, 14 strains of Ralstonia pickettii were isolated from various biological samples inoculated in a blood culture medium. Molecular epidemiological investigation confirmed the relatedness of the strains. The source of the contamination proved to be the blood culture bottle caps. PMID:12089303

  9. Inhibition of Salmonella Typhimurium by Cultures of Cecal Bacteria during Aerobic Incubation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two trials were conducted to examine the ability of cecal bacterial cultures from broilers to inhibit growth of Salmonella Typhimurium during aerobic incubation. Cecal broth media was inoculated with 10 µl of cecal contents from 6 week old broilers taken from 2 separate flocks. Cultures were incubat...

  10. Effect of 12-week-long aerobic training programme on body composition, aerobic capacity, complete blood count and blood lipid profile among young women

    PubMed Central

    Nowak, Robert; Jastrzębski, Zbigniew; Zarębska, Aleksandra; Bichowska, Marta; Drobnik-Kozakiewicz, Izabela; Radzimiński, Łukasz; Leońska-Duniec, Agata; Ficek, Krzysztof; Cięszczyk, Paweł

    2015-01-01

    Background Numerous data suggest that aerobic-type exercise improves lipoprotein-lipid profiles, cardiorespiratory fitness and body composition in young women. The aim of this study was to evaluate the biological response to high-low impact aerobic fitness among young women. Materials and methods Thirty-four young women aged 22 (19-24) years were divided into three groups: underweight (N = 10), normal weight (N = 12) and overweight (N = 12). Aerobic capacity, anthropometry and body composition together with complete blood count and lipid profile were determined before and after completion of a 12-week-long training period. Results The training programme caused a significant decrease in weight (by 4.3 kg, P = 0.003), body mass index (by 1.3 kg/m2, P = 0.003), free fat mass (by 2.1 kg, P = 0.002), total body water (by 0.4 kg, P = 0.036), percentage of fat (by 3 percent points, P = 0.002), all analyzed skinfolds thicknesses, as well as the lipid profile in overweight group, and no changes in normal weight group. Significant changes in weight (by 4.2 kg, P = 0.005), body mass index (by 0.9 kg/m2, P = 0.005), crus skinfold thickness (by 3.3 mm, P = 0.028), and in maximum oxygen uptake (by 2.49 mL/kg/min; P = 0.047) were observed among underweight women. No change in total blood count was observed in all groups. Conclusion Twelve-week-long fitness training programme of two alternating styles (low and high impact) has a beneficial effect on overweight young women. PMID:25672474

  11. Use of a pair of blood culture bottles for sterility testing of corneal organ culture media

    PubMed Central

    Gain, P.; Thuret, G.; Chiquet, C.; Vautrin, A.; Carricajo, A.; Acquart, S.; Maugery, J.; Aubert, G.

    2001-01-01

    AIMS—To test the effectiveness and rapidity of a pair of blood culture bottles in the diagnosis of bacterial and fungal contamination of corneal organ culture media.
METHODS—761 microbiological analyses of storage media (Inosol and Exosol, Opsia, Toulouse, France), sampled in all phases of the organ culture at 31°C of 410 consecutive corneas, were analysed. Each medium was inoculated in a pair of Bactec Plus Aerobic/F and Bactec Lytic/10 Anaerobic/F blood bottles and placed in a Bactec 9240 incubator for 14 days at 37°C and in a Sabouraud broth at 20°C. Changes in colour or turbidity of storage media were evaluated daily at the corneal bank. Recipients were screened post-graft for infectious signs.
RESULTS—Overall contamination rate was 2.4% (18/761). Contamination was detected in less than 1 day in 78% (14/18) and less than 2 days in 94% (17/18). Positivity of the microbiological controls of starting media preceded changes medium colour in 10 out of 14 cases. Bactec blood bottles allowed detection of bacteria as well as yeasts.
CONCLUSION—The use of a pair of Bactec blood culture bottles appears reliable for the rapid diagnosis of a wide range of microbiological contaminations of organ cultured corneas during banking.

 PMID:11567956

  12. Aerobic power and the main determinants of blood rheology: is there a relationship?

    PubMed

    El-Sayed, Mahmoud S; Ali, Nagia; Al-Bayatti, Mudhaffar

    2009-12-01

    This cross-sectional study aimed to explore the relationship between aerobic power and the main determinants of blood rheology namely plasma viscosity, plasma fibrinogen concentration and haematocrit. Ninety-three normal healthy individuals (VO2max 48.3 ml/kg per min), who were familiarized with the laboratory environment and testing procedures, participated in the study. Aerobic power as assessed by maximal oxygen consumption (VO2max) was determined by using an incremental exercise protocol on either a treadmill or a stationary bike. Oxygen consumption was measured online using a computer-based metabolic system. In a standardized resting condition, venous blood samples were removed from a prominent vein of the nondominant arm. Aliquots of whole blood were measured for haematocrit (in triplicate), whereas plasma was assayed for fibrinogen concentration and viscosity (in duplicate) using semiautomatic coagulometer and capillary viscometer; respectively. The mean values for haematocrit (41.9 +/- 2.5%), plasma viscosity (1.56 +/- 0.27 mPa s) and plasma fibrinogen (272.1 +/- 86.9 mg/dl) were within the normal range for normal participants. Pearson correlation coefficients and regression analysis were used for statistical evaluations. In this population, VO2max negatively correlated with plasma viscosity (P < 0.01) and plasma fibrinogen concentration (P < 0.01). Although VO2max positively correlated with haematocrit, this correlation was not as strong. Thus, high aerobic power as assessed by maximal oxygen consumption appears to be associated with lower plasma viscosity and lower plasma fibrinogen. The significant negative relationships between VO2max and plasma viscosity and plasma fibrinogen might suggest that blood is more dilute in individuals with high aerobic power. This could probably be due to an expansion of plasma volume, which is commonly seen in those who are physically active and exhibit a higher level of cardiorespiratory fitness. PMID:19786866

  13. Clinical implications of positive blood cultures.

    PubMed Central

    Bryan, C S

    1989-01-01

    Positive blood cultures can be classified according to their veracity (true-positive or false-positive culture), clinical severity (inconsequential or life threatening), place of origin (community acquired or nosocomial), source (primary or secondary), duration (transient, intermittent, or continuous), pattern of occurrence (single episode, persistent, or recurrent), or intensity (high or low grade). In general, however, positive blood cultures identify a patient population at high risk of death. In my studies, patients with positive blood cultures were 12 times more likely to die during hospitalization than patients without positive blood cultures. Many bacteremias and fungemias occur in complicated clinical settings, and it appears that only about one-half of the deaths among affected patients are due directly to infection. Hence, it is appropriate to speak of "crude mortality" and "attributable mortality." Among hospitalized patients, recent trends include rising incidences of Staphylococcus aureus and coagulase-negative staphylococcal and enterococcal bacteremias and a dramatic increase in the incidence of fungemias. The diagnostic and therapeutic implications of blood cultures positive for specific microorganisms continue to evolve and are the subject of a large and growing medical literature. PMID:2680055

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

    PubMed Central

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

    2016-01-01

    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

  15. Simplified lysed-blood culture technique.

    PubMed Central

    Zierdt, C H

    1986-01-01

    A blood culture system was developed in which a lysing agent (either Tween 20, one of several other polyoxyethylene adducts, digitonin, or Triton X-100) is added to the blood culture medium. Of 33 Triton compounds, 9 lysed human blood, as did 7 of 21 polyoxyethylene compounds and digitonin, all at a concentration of 0.05%. Under the specific test conditions, three of the hemolytic polyoxyethylene compounds and digitonin had no inhibitory effect. All of the Triton compounds had at least some inhibitory effect on the most sensitive of the pathogenic bacteria that were tested, Streptococcus pneumoniae and Neisseria meningitidis. Because of results from previous studies, Triton X-100 was tested further, despite evidence in this study of its inhibition of bacteria. Of the 55 lysing agents tested, digitonin, Triton X-100, Brij 96, and Tween 20 were selected for further testing as additions to conventional culture broth. Comparative culture studies with bacteremic blood from infected rabbits were performed with the conventional blood culture, the Isolator system (Du Pont Co., Wilmington, Del.), and the new lysing medium. The new system has the advantages of lysis filtration and lysis centrifugation without the associated added cost and processing complexity. PMID:3958142

  16. Rapid presumptive identification of anaerobes in blood cultures by gas-liquid chromatography.

    PubMed Central

    Sondag, J E; Ali, M; Murray, P R

    1980-01-01

    Production of volatile and nonvolatile metabolic acids in blood culture broths by aerobic, facultative anaerobic, and obligate anaerobic bacteria was analyzed by gas-liquid chromatography. Anaerobic blood culture isolates were presumptively identified by the qualitative analysis of volatile fatty acids. Isolates, with a characteristic Gram stain reaction and cellular morphology, were identified by the following acid patterns: Bacteriodes fragilis group with acetic and propionic acids; Fusobacterium with acetic, butyric, and usually propionic acids; Veillonella with acetic and propionic acids; gram-positive cocci with acetic and butyric acids; and Clostridium with acetic and butyric acids. PMID:7381002

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

    PubMed Central

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

    2016-01-01

    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. PMID:26904669

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

    PubMed

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

    1984-10-01

    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

  19. Flow microcalorimetry investigation of the influence of surfactants on a heterogeneous aerobic culture.

    PubMed Central

    Beaubien, A; Keita, L; Jolicoeur, C

    1987-01-01

    The influence of various surfactants on the biological activity of a mixed aerobic culture has been investigated by using flow microcalorimetry. The response of the culture to the addition of homologous n-alkylcarboxylates (C2 to C16) and n-alkylpyridinium bromides (C11 to C14) has been examined under endogenous and substrate saturation conditions, and inhibitory concentrations (MIC or the concentration which decreased the initial activity (heat flux) of the culture by 50%) were determined for each state. Under both conditions, the n-alkylpyridinium bromides were found to be more toxic than the n-alkylcarboxylates of identical chain length, thus confirming that the head group of the amphiphiles plays an important role in the microbial toxicity of surfactants. The relationship observed between the concentration at which 50% of the activity is lost and the chain length of the surfactant further confirms that cellular toxicity is also dependent on surfactant hydrophobicity. In relation to the biodegradability of surfactants in mixed aerobic cultures, the low concentration effects of n-alkylcarboxylates on endogenous culture were investigated in some detail. There appear to be compounded indications that these surfactants are rapidly metabolized by the microorganisms of the mixed culture, at least for homologs lower than C10. PMID:3426221

  20. Evaluation of nitrate removal by continuous culturing of an aerobic denitrifying bacterium, Paracoccus pantotrophus.

    PubMed

    Hasegawa-Kurisu, K; Otani, Y; Hanaki, K

    2006-01-01

    Nitrate removal under aerobic conditions was investigated using pure cultures of Paracoccus pantotrophus, which is a well-known aerobic-denitrifying (AD) bacterium. When a high concentration of cultures with a high carbon/nitrogen (C/N) ratio was preserved at the beginning of batch experiments, subsequently added nitrate was completely removed. When continuous culturing was perpetuated, a high nitrate removal rate (66.5%) was observed on day 4 post-culture, although gradual decreases in AD ability with time were observed. The attenuation in AD ability was probably caused by carbon limitation, because when carbon concentration of inflow water was doubled, nitrate removal efficiency improved from 18.1% to 59.6%. Bacterial community analysis using the polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) method showed that P. pantotrophus disappeared in the suspended medium on day 8 post-culture, whereas other bacterial communities dominated by Acidovorax sp. appeared. Interestingly, this replaced bacterial community also showed AD ability. As P. pantotrophus was detected as attached colonies around the membrane and bottom of the reactor, this bacterium can therefore be introduced in a fixed form for treatment of wastewater containing nitrate with a high C/N ratio. PMID:17163031

  1. Aerobic culture of methanogenic archaea without an external source of hydrogen.

    PubMed

    Khelaifia, S; Lagier, J-C; Nkamga, V D; Guilhot, E; Drancourt, M; Raoult, D

    2016-06-01

    Culturing methanogenic archaea is fastidious, expensive, and requires an external source of hydrogen and carbon dioxide. Until now, these microorganisms have only been cultivated under strictly anaerobic conditions. We previously developed a single versatile culture medium containing sugars and anti-oxydants for cultivating all human known methanogens. Performing aerobic cultures in the presence of Bacteroides thetaiotaomicron, which produces hydrogen, allows for cultivation of Methanobrevibacter smithii which itself produces methane. To obtain colonies, we cultivated M. smithii in an agar plate in the upper part of a double chamber flask with a liquid culture of B. thetaiotaomicron in the lower compartment. We subsequently cultured four other methanogenic species for the first time and successfully isolated 13 strains of M. smithii and nine strains of Methanobrevibacter oralis from 100 stools and 45 oral samples. This procedure allows aerobic isolation and antibiotic susceptibility testing. This changes the ability to routinely study methanogens, which have been neglected in clinical microbiology laboratories and may be useful for biogas production. PMID:27010812

  2. Comparison of the Roche Septi-Chek blood culture bottle with a brain heart infusion biphasic medium bottle and with a tryptic soy broth bottle.

    PubMed Central

    Henry, N K; Grewell, C M; McLimans, C A; Washington, J A

    1984-01-01

    In a comparison of 1,368 positive blood cultures, a vented Roche Septi-Chek (V-RSC) blood culture bottle was superior to an unvented tryptic soy broth-containing bottle (Difco) for the recovery of all aerobic and facultatively anaerobic microorganisms. Anaerobic bacteria were recovered more frequently and earlier in the unvented tryptic soy broth-containing bottle. A separate comparison of 529 positive blood cultures was conducted to examine the performance of the V-RSC bottle with that of a vented brain heart infusion biphasic medium. The V-RSC bottle recovered significantly more isolates of Enterobacteriaceae and of anaerobic bacteria than did the vented brain heart infusion biphasic medium. The V-RSC bottle is a reliable blood culture system for all aerobic and facultatively anaerobic microorganisms. Because of its suboptimal recovery of anaerobic bacteria, it is recommended that the V-RSC bottle be used in combination with an unvented vacuum blood culture bottle. PMID:6371039

  3. The Effects of 8-Weeks Aerobic Exercise Program on Blood Lipids and Cholesterol Profile of Smokers vs. Non Smokers

    ERIC Educational Resources Information Center

    Taifour, Akef; AL-Shishani, Ahmad; Khasawneh, Aman; AL-Nawaiseh, Ali; Bakeer, Mohammed

    2015-01-01

    The aim of this study was to compare the effects of 8-week aerobic exercise program on blood lipids and cholesterol profile of smoker's vs. non-smokers. A total of 34 male subjects (18 non-smokers and 16 smokers) took part in this study. Both groups were pre- and post tested in their blood-lipids and cholesterol profile before and after the 8-week…

  4. Storage of Stock Cultures of Filamentous Fungi, Yeasts, and Some Aerobic Actinomycetes in Sterile Distilled Water

    PubMed Central

    McGinnis, M. R.; Padhye, A. A.; Ajello, L.

    1974-01-01

    Castellani's procedure for maintaining cultures of filamentous fungi and yeasts in sterile distilled water was evaluated. Four hundred and seventeen isolates of 147 species belonging to 66 genera of filamentous fungi, yeasts, and aerobic actinomycetes were maintained in sterile distilled water at room temperature over periods ranging from 12 to 60 months in four independent experiments. Of the 417 cultures, 389 (93%) survived storage in sterile distilled water. The selection of good sporulating cultures and sufficient inoculum consisting of spores and hyphae suspended in sterile distilled water were the most important factors influencing survival in water over a longer period of time. The technique was found to be simple, inexpensive, and reliable. PMID:4854418

  5. Batch culture enrichment of ANAMMOX populations from anaerobic and aerobic seed cultures.

    PubMed

    Suneethi, S; Joseph, Kurian

    2011-01-01

    Discharge of nitrate and ammonia rich wastewaters into the natural waters encourage eutrophication, and contribute to aquatic toxicity. Anaerobic ammonium oxidation process (ANAMMOX) is a novel biological nitrogen removal alternative to nitrification-denitrification, that removes ammonia using nitrite as the electron acceptor. The feasibility of enriching the ANAMMOX bacteria from the anaerobic digester sludge of a biomethanation plant treating vegetable waste and aerobic sludge from an activated sludge process treating domestic sewage is reported in this paper. ANAMMOX bacterial activity was monitored and established in terms of nitrogen transformations to ammonia, nitrite and nitrate along with formation of hydrazine and hydroxylamine. PMID:20729077

  6. Controlled aerobic exercise training reduces resting blood pressure in sedentary older adults.

    PubMed

    Huang, Guoyuan; Shi, Xiangrong; Gibson, Cheryl A; Huang, Sunny C; Coudret, Nadine A; Ehlman, Mary C

    2013-12-01

    The results of existing controlled clinical trials were synthesized to determine effects of aerobic exercise training on resting systolic (SBP) and diastolic blood pressure (DBP) among previously sedentary older adults, to quantify the magnitude of observed changes, and to examine the influence of the associated interventional variables on these changes. Studies were identified via a systematic computer database search, hand searching, and cross-referencing of previously located articles. All potentially eligible articles were carefully reviewed and examined with the established inclusion criteria. Twenty-three studies, representing a total of 1226 older subjects, were included in the final analysis. Robust statistically significant effects were found in terms of the pooled standardized effect size of - 0.33 ± 0.06 (p < 0.0001) in SBP and - 0.39 ± 0.09 (p < 0.0001) in DBP. When compared with the control group, net decreases in both SBP (- 5.39 ± 1.21 mmHg, p < 0.0001) and DBP (-3.68 ± 0.83 mmHg, p < 0.0001) were observed in older exercisers, representing a 3.9% and a 4.5% reduction, respectively. This meta-analytic study provides robust quantitative data to support the efficacy and effectiveness of controlled endurance exercise training in decreasing resting SBP and DBP among previously sedentary older adults. PMID:23550511

  7. Six weeks of aerobic dance exercise improves blood oxidative stress status and increases interleukin-2 in previously sedentary women.

    PubMed

    Leelarungrayub, Donrawee; Saidee, Kunteera; Pothongsunun, Prapas; Pratanaphon, Sainetee; YanKai, Araya; Bloomer, Richard J

    2011-07-01

    This study evaluated the change in blood oxidative stress, blood interleukin-2, and physical performance following 6 weeks of moderate intensity and duration aerobic dance exercise in 24 sedentary women. Blood samples were collected at rest twice before (baseline) and after the 6-week intervention for analysis of protein hydroperoxide (PrOOH), malondialdehyde (MDA), total anti-oxidant capacity (TAC), and interleukin-2 (IL-2) levels. Maximal treadmill run time (Time(max)) and maximal oxygen consumption (VO(2max)) were also measured. All variables were statistically analyzed with a repeated measurement ANOVA and Tukey post hoc. No differences were noted in any variable during the baseline period (p > 0.05). After aerobic dance exercise, VO(2max), Time(max), TAC and IL-2 were significantly increased, whereas MDA levels were decreased significantly (p < 0.05). PrOOH did not change either between baseline measures or after exercise. It can be concluded that aerobic dance exercise at a moderate intensity and duration can improve physical fitness, decrease MDA, and increase TAC and IL-2 in previously sedentary women. PMID:21665113

  8. Long-term storage of aerobic granules in liquid media: viable but non-culturable status.

    PubMed

    Wan, Chunli; Zhang, Qinlan; Lee, Duu-Jong; Wang, Yayi; Li, Jieni

    2014-08-01

    Long-term storage and successful reactivation after storage are essential for practical applications of aerobic granules on wastewater treatment. This study cultivated aerobic granules (SI) in sequencing batch reactors and then stored the granules at 4 °C in five liquid media (DI water (SW), acetone (SA), acetone/isoamyl acetate mix (SAA), saline water (SS), and formaldehyde (SF)) for over 1 year. The first four granules were then successfully reactivated in 24h cultivation. The specific oxygen uptake rates (SOUR) of the granules followed SI>SS>SA>SAA>SW>SF; and the corresponding granular strengths (10 min ultrasound) followed SI>SA=SS>SAA>SW>SF. During storage the granular cells secreted excess quantities of cyclic-diguanylate (c-di-GMP) and pentaphosphate (ppGpp) as responses to the stringent challenges. We proposed that to force cells in granules (Alphaproteobacteria, Flavobacteria, Betaproteobacteria, Gammaproteobacteria, Actinobacteria, Sphingobacteria, and Clostridia) entering viable but non-culturable (VBNC) status is the key of success for extended period storage of granules. PMID:24950091

  9. Direct Isolation of Candida spp. from Blood Cultures on the Chromogenic Medium CHROMagar Candida

    PubMed Central

    Horvath, Lynn L.; Hospenthal, Duane R.; Murray, Clinton K.; Dooley, David P.

    2003-01-01

    CHROMagar Candida is a selective and differential chromogenic medium that has been shown to be useful for identification of Candida albicans, Candida krusei, Candida tropicalis, and perhaps Candida glabrata. Colony morphology and color have been well defined when CHROMagar Candida has been used to isolate yeast directly from clinical specimens, including stool, urine, respiratory, vaginal, oropharyngeal, and esophageal sources. Direct isolation of yeast on CHROMagar Candida from blood cultures has not been evaluated. We evaluated whether the color and colony characteristics produced by Candida spp. on CHROMagar Candida were altered when yeasts were isolated directly from blood cultures. Fifty clinical isolates of Candida were inoculated into aerobic and anaerobic blood culture bottles and incubated at 35°C in an automated blood culture system. When growth was detected, an aliquot was removed and plated onto CHROMagar Candida. As a control, CHROMagar Candida plates were inoculated with the same isolate of yeast grown on Sabouraud dextrose agar simultaneously. No significant difference was detected in color or colony morphology between the blood and control isolates in any of the tested organisms. All C. albicans (n = 12), C. tropicalis (n = 12), C. glabrata (n = 9), and C. krusei (n = 5) isolates exhibited the expected species-specific colony characteristics and color, whether isolated directly from blood or from control cultures. CHROMagar Candida can be reliably used for direct isolation of yeast from blood cultures. Direct isolation could allow mycology laboratories to more rapidly identify Candida spp., enable clinicians to more quickly make antifungal agent selections, and potentially decrease patient morbidity and mortality. PMID:12791890

  10. Effect of selected monoterpenes on methane oxidation, denitrification, and aerobic metabolism by bacteria in pure culture.

    PubMed

    Amaral, J A; Ekins, A; Richards, S R; Knowles, R

    1998-02-01

    Selected monoterpenes inhibited methane oxidation by methanotrophs (Methylosinus trichosporium OB3b, Methylobacter luteus), denitrification by environmental isolates, and aerobic metabolism by several heterotrophic pure cultures. Inhibition occurred to various extents and was transient. Complete inhibition of methane oxidation by Methylosinus trichosporium OB3b with 1.1 mM (-)-alpha-pinene lasted for more than 2 days with a culture of optical density of 0.05 before activity resumed. Inhibition was greater under conditions under which particulate methane monooxygenase was expressed. No apparent consumption or conversion of monoterpenes by methanotrophs was detected by gas chromatography, and the reason that transient inhibition occurs is not clear. Aerobic metabolism by several heterotrophs was much less sensitive than methanotrophy was; Escherichia coli (optical density, 0.01), for example, was not affected by up to 7.3 mM (-)-alpha-pinene. The degree of inhibition was monoterpene and species dependent. Denitrification by isolates from a polluted sediment was not inhibited by 3.7 mM (-)-alpha-pinene, gamma-terpinene, or beta-myrcene, whereas 50 to 100% inhibition was observed for isolates from a temperate swamp soil. The inhibitory effect of monoterpenes on methane oxidation was greatest with unsaturated, cyclic hydrocarbon forms [e.g., (-)-alpha-pinene, (S)-(-)-limonene, (R)-(+)-limonene, and gamma-terpinene]. Lower levels of inhibition occurred with oxide and alcohol derivatives [(R)-(+)-limonene oxide, alpha-pinene oxide, linalool, alpha-terpineol] and a noncyclic hydrocarbon (beta-myrcene). Isomers of pinene inhibited activity to different extents. Given their natural sources, monoterpenes may be significant factors affecting bacterial activities in nature. PMID:9464387

  11. Coordination of Metabolism and Virulence Factors Expression of Extraintestinal Pathogenic Escherichia coli Purified from Blood Cultures of Patients with Sepsis.

    PubMed

    Pettersen, Veronika Kuchařová; Mosevoll, Knut Anders; Lindemann, Paul Christoffer; Wiker, Harald G

    2016-09-01

    One of the trademarks of extraintestinal pathogenic Escherichia coli is adaptation of metabolism and basic physiology to diverse host sites. However, little is known how this common human pathogen adapts to permit survival and growth in blood. We used label-free quantitative proteomics to characterize five E. coli strains purified from clinical blood cultures associated with sepsis and urinary tract infections. Further comparison of proteome profiles of the clinical strains and a reference uropathogenic E. coli strain 536 cultivated in blood culture and on two different solid media distinguished cellular features altered in response to the pathogenically relevant condition. The analysis covered nearly 60% of the strains predicted proteomes, and included quantitative description based on label-free intensity scores for 90% of the detected proteins. Statistical comparison of anaerobic and aerobic blood cultures revealed 32 differentially expressed proteins (1.5% of the shared proteins), mostly associated with acquisition and utilization of metal ions critical for anaerobic or aerobic respiration. Analysis of variance identified significantly altered amounts of 47 proteins shared by the strains (2.7%), including proteins involved in vitamin B6 metabolism and virulence. Although the proteomes derived from blood cultures were fairly similar for the investigated strains, quantitative proteomic comparison to the growth on solid media identified 200 proteins with substantially changed levels (11% of the shared proteins). Blood culture was characterized by up-regulation of anaerobic fermentative metabolism and multiple virulence traits, including cell motility and iron acquisition. In a response to the growth on solid media there were increased levels of proteins functional in aerobic respiration, catabolism of medium-specific carbon sources and protection against oxidative and osmotic stresses. These results demonstrate on the expressed proteome level that expression of

  12. An initial investigation into the ecology of culturable aerobic postmortem bacteria.

    PubMed

    Chun, Lauren P; Miguel, Marcus J; Junkins, Emily N; Forbes, Shari L; Carter, David O

    2015-12-01

    Postmortem microorganisms are increasingly recognized for their potential to serve as physical evidence. Yet, we still understand little about the ecology of postmortem microbes, particularly those associated with the skin and larval masses. We conducted an experiment to characterize microbiological and chemical properties of decomposing swine (Sus scrofa domesticus) carcasses on the island of Oahu, Hawaii, USA, during June 2013. Bacteria were collected from the head, limb, and larval mass during the initial 145h of decomposition. We also measured the pH, temperature, and oxidation-reduction potential of larval masses in situ. Bacteria were cultured aerobically on Standard Nutrient Agar at 22°C and identified using protein or genetic signals. Carcass decomposition followed a typical sigmoidal pattern and associated bacterial communities differed by sampling location and time since death, although all communities were dominated by phyla Actinobacteria, Firmicutes, and Proteobacteria. Larval masses were reducing environments (~-200mV) of neutral pH (6.5-7.5) and high temperature (35°C-40°C). We recommend that culturable postmortem and larval mass microbiology and chemistry be investigated in more detail, as it has potential to complement culture-independent studies and serve as a rapid estimate of PMI. PMID:26654073

  13. Robustness of an aerobic metabolically vinyl chloride degrading bacterial enrichment culture.

    PubMed

    Zhao, He-Ping; Schmidt, Kathrin R; Lohner, Svenja; Tiehm, Andreas

    2011-01-01

    Degradation of the lower chlorinated ethenes is crucial to the application of natural attenuation or in situ bioremediation on chlorinated ethene contaminated sites. Recently, within mixtures of several chloroethenes as they can occur in contaminated groundwater inhibiting effects on aerobic chloroethene degradation have been shown. The current study demonstrated that metabolic vinyl chloride (VC) degradation by an enrichment culture originating from groundwater was not affected by an equimolar concentration (50 μM) of cis-1,2-dichloroethene (cDCE). Only cDCE concentrations at a ratio of 2.4:1 (initial cDCE to VC concentration) caused minor inhibition of VC degradation. Furthermore, the degradation of VC was not affected by the presence of trans-1,2-dichloroethene (tDCE), 1,1-dichloroethene (1,1-DCE), trichloroethene (TCE), and tetrachloroethene (PCE) in equimolar concentrations (50 μM). Only cDCE and tDCE were cometabolically degraded in small amounts. The VC-degrading culture demonstrated a broad pH tolerance from 5 to 9 with an optimum between 6 and 7. Results also showed that the culture could degrade VC concentrations up to 1,800 μM (110 mg/L). PMID:22020471

  14. Isolation of culturable aerobic bacteria and evidence of Kerstersia gyiorum from the blowhole of captive Yangtze finless porpoises.

    PubMed

    Wan, Xiaoling; McLaughlin, Richard William; Zhou, Junying; Hao, Yujiang; Zheng, Jinsong; Wang, Ding

    2016-08-01

    Bacterial respiratory illnesses are problematic in aquatic mammals such as the Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis; YFP), which is now at a critically endangered status. Yet little is known about the bacteria inhabiting the respiratory tract of YFPs. In this study, we preliminarily characterized the culturable aerobic bacteria from blow samples of captive YFPs. The bacterial diversity was assessed through cultivation by direct exhalation onto Columbia blood agar plates and identification of representative isolates through 16S rRNA gene sequence analysis. In total, eleven bacterial species belonging to four phyla Proteobacteria (71 %), Firmicutes (25 %), Bacteroidetes (3 %) and Actinobacteria (1 %) were identified. Most of these isolates were opportunistic pathogens found in respiratory illnesses in humans and animals. We also reported the first case of Kerstersia gyiorum isolated from an animal. This work provides a preliminary assessment of the bacteria present in the respiratory tract of captive YFPs, which will be an important first step in elucidating the roles of normal microbiota in maintaining respiratory health of YFPs. This study also points out the necessity of future long-term monitoring of blowhole microorganisms in the YFPs and making emergency preparedness plans for respiratory tract infections. These measures can aid in assessing the pathogenic risk of the critically endangered YFP populations. PMID:27251558

  15. Water aerobics is followed by short-time and immediate systolic blood pressure reduction in overweight and obese hypertensive women.

    PubMed

    Cunha, Raphael Martins; Arsa, Gisela; Neves, Eduardo Borba; Lopes, Lorena Curado; Santana, Fabio; Noleto, Marcelo Vasconcelos; Rolim, Thais I; Lehnen, Alexandre Machado

    2016-07-01

    One exercise training session such as walking, running, and resistance can lead to a decrease in blood pressure in normotensive and hypertensive individuals, but few studies have investigated the effects of exercise training in an aquatic environment for overweight and obese hypertensive individuals. We aimed to assess the acute effects of a water aerobics session on blood pressure changes in pharmacologically treated overweight and obese hypertensive women. A randomized crossover study was carried out with 18 hypertensive women, 10 of them were overweight (54.4 ± 7.9 years; body mass index: 27.8 ± 1.7 kg/m(2)) and eight obese (56.4 ± 6.6 years; body mass index: 33.0 ± 2.0 kg/m(2)). The water aerobics exercise session consisted of a 45-minute training at the intensity of 70%-75% of maximum heart rate adjusted for the aquatic environment. The control group did not enter the pool and did not perform any exercise. We measured systolic blood pressure (SBP) and diastolic blood pressure (DBP) before, immediately after, and every 10 minutes up to 30 minutes after the aerobic exercise or control session. Overall (n = 18), DBP did not change after the water aerobic exercise and control session, and SBP decreased at 10 and 20 minutes postexercise compared to the control session. Among overweight women, SBP decreased at 10 and 20 minutes postexercise. In contrast, among obese women, SBP decreased only at 10 minutes postexercise. SBP variation was -2.68 mm Hg in overweight and -1.24 mm Hg in obese women. In conclusion, the water aerobics session leads to a reduction in SBP, but not in DBP, during 10 and 20 minutes postexercise recovery. Thus, it may be safely prescribed to overweight and obese women. PMID:27245928

  16. Respiration and respiratory enzyme activity in aerobic and anaerobic cultures of the marine denitrifying bacterium, Pseudomonas perfectomarinus

    NASA Astrophysics Data System (ADS)

    Packard, T. T.; Garfield, P. C.; Martinez, R.

    1983-03-01

    Oxygen consumption, nitrate reduction, respiratory electron transport activity, and nitrate reductase activity were measured in aerobic and anaerobic cultures of the marine bacterium, Pseudomonas perfectomarinus. The respiratory electron transport activity was closely correlated with oxygen consumption ( r = 0.98) in aerobic cultures and nearly as well correlated with nitrate reductase activity ( r = 0.91) and nitrate reduction ( r = 0.85) in anaerobic cultures. It was also well correlated with biomass in both aerobic ( r = 0.99) and anaerobic ( r = 0.94) cultures supporting the use of tetrazolium reduction as an index of living biomass. Time courses of nitrate and nitrate in the anaerobic cultures demonstrated that at nitrate concentrations above 1 mM, denitrification proceeds stepwise. Time courses of pH in anaerobic cultures revealed a rise from 7 to 8.5 during nitrite reduction indicating net proton utilization. This proton utilization is predicted by the stoichiometry of denitrification. Although the experiments were not under 'simulated in situ' conditions, the results are relevant to studies of denitrification, to bacterial ATP production, and to the respiratory activity of marine plankton in the ocean.

  17. Benzoate-induced stress enhances xylitol yield in aerobic fed-batch culture of Candida mogii TISTR 5892.

    PubMed

    Wannawilai, Siwaporn; Sirisansaneeyakul, Sarote; Chisti, Yusuf

    2015-01-20

    Production of the natural sweetener xylitol from xylose via the yeast Candida mogii TISTR 5892 was compared with and without the growth inhibitor sodium benzoate in the culture medium. Sodium benzoate proved to be an uncompetitive inhibitor in relatively poorly oxygenated shake flask aerobic cultures. In a better controlled aerobic environment of a bioreactor, the role of sodium benzoate could equally well be described as competitive, uncompetitive or noncompetitive inhibitor of growth. In intermittent fed-batch fermentations under highly aerobic conditions, the presence of sodium benzoate at 0.15gL(-1) clearly enhanced the xylitol titer relative to the control culture without the sodium benzoate. The final xylitol concentration and the average xylitol yield on xylose were nearly 50gL(-1) and 0.57gg(-1), respectively, in the presence of sodium benzoate. Both these values were substantially higher than reported for the same fermentation under microaerobic conditions. Therefore, a fed-batch aerobic fermentation in the presence of sodium benzoate is promising for xylitol production using C. mogii. PMID:25499077

  18. Evaluation of the FilmArray Blood Culture Identification Panel: Results of a Multicenter Controlled Trial

    PubMed Central

    Salimnia, Hossein; Lephart, Paul R.; Schreckenberger, Paul; DesJarlais, Sharon M.; Johnson, J. Kristie; Robinson, Gwen; Carroll, Karen C.; Greer, Amy; Morgan, Margie; Chan, Raymond; Loeffelholz, Michael; Valencia-Shelton, Frances; Jenkins, Stephen; Schuetz, Audrey N.; Daly, Judy A.; Barney, Trenda; Hemmert, Andrew; Kanack, Kristen J.

    2016-01-01

    Sepsis is a major cause of morbidity, mortality, and increased medical expense. Rapid diagnosis improves outcomes and reduces costs. The FilmArray blood culture identification panel (BioFire Diagnostics LLC, Salt Lake City, UT), a highly multiplexed PCR assay, can identify 24 etiologic agents of sepsis (8 Gram-positive, 11 Gram-negative, and 5 yeast species) and three antimicrobial resistance genes (mecA, vanA/B, and blaKPC) from positive blood culture bottles. It provides results in about 1 h with 2 min for assay setup. We present the results of an eight-center trial comparing the sensitivity and specificity of the panel with those of the laboratories' standard phenotypic identification techniques, as well as with molecular methods used to distinguish Acinetobacter baumannii from other members of the A. calcoaceticus-A. baumannii complex and to detect antimicrobial resistance genes. Testing included 2,207 positive aerobic blood culture samples, 1,568 clinical and 639 seeded. Samples were tested fresh or were frozen for later testing within 8 h after the bottles were flagged as positive by an automated blood culture system. At least one organism was detected by the panel in 1,382 (88.1%) of the positive clinical specimens. The others contained primarily off-panel organisms. The panel reported multiple organisms in 81 (5.86%) positive clinical specimens. The unresolved blood culture identification sensitivity for all target detections exceeded 96%, except for Klebsiella oxytoca (92.2%), which achieved 98.3% sensitivity after resolution of an unavoidable phenotypic error. The sensitivity and specificity for vanA/B and blaKPC were 100%; those for mecA were 98.4 and 98.3%, respectively. PMID:26739158

  19. Stoichiometry and kinetics of poly-{beta}-hydroxybutyrate metabolism in aerobic, slow growing, activated sludge cultures

    SciTech Connect

    Beun, J.J.; Paletta, F.; Loosdrecht, M.C.M. Van; Heijnen, J.J.

    2000-02-20

    This paper discusses the poly-{beta}-hydroxybutyrate (PHB) metabolism in aerobic, slow growing, activated sludge cultures, based on experimental data and on a metabolic model. The dynamic conditions which occur in activated sludge processes were simulated in a 2-L sequencing batch reactor (SBR) by subjecting a mixed microbial population to successive periods of external substrate availability (feast period) and no external substrate availability (famine period). Under these conditions intracellular storage and consumption of PHB was observed. It appeared that in the feast period, 66% to almost 100% of the substrate consumed is used for storage of PHB, the remainder is used for growth and maintenance processes. Furthermore, it appeared that at high sludge retention time (SRT) the growth rate in the feast and famine periods was the same. With decreasing SRT the growth rate in the feast period increased relative to the growth rate in the famine period. Acetate consumption and PHB production in the feast period both proceeded with a zero-order rate in acetate and PHB concentration respectively. PHB consumption in the famine period could best be described kinetically with a nth order degradation equation in PHB concentration. The obtained results are discussed in the context of the general activated sludge models.

  20. High prevalence of Kingella kingae in joint fluid from children with septic arthritis revealed by the BACTEC blood culture system.

    PubMed Central

    Yagupsky, P; Dagan, R; Howard, C W; Einhorn, M; Kassis, I; Simu, A

    1992-01-01

    In an effort to improve detection of fastidious organisms, joint fluid aspirates of pediatric patients were inoculated into BACTEC 460 aerobic blood culture bottles, in addition to cultures on solid media. Culture records for the 1988 to 1991 period were reviewed to compare the performance of both methods for the recovery of pathogens. Overall, 216 children underwent a diagnostic joint tap, and 63 specimens grew significant organisms, including Kingella kingae in 14. While both methods were comparable for recovery of usual pathogens, with a single exception, K. kingae isolates were detected by the BACTEC system only. K. kingae appears to be a more common cause of septic arthritis in children than has been previously recognized. The BACTEC blood culture system enhances the recovery of K. kingae from joint fluid and improves bacteriologic diagnosis of pediatric septic arthritis. PMID:1583131

  1. Aerobic Exercise Attenuates an Exaggerated Exercise Blood Pressure Response in Normotensive Young Adult African-American Men

    PubMed Central

    BOND, VERNON; STEPHENS, QUIONA; ADAMS, RICHARD G.; VACCARO, PAUL; DEMEERSMAN, RONALD; WILLIAMS, DEBORAH; OBISESAN, THOMAS O.; FRANKS, B. DON; OKE, LUE M.; COLEMAN, BERNELL; BLAKELY, RAYMOND; MILLIS, RICHARD M.

    2011-01-01

    An exaggerated exercise blood pressure response (EEBPR) may be associated with an increased risk of hypertension. We hypothesized that aerobic exercise training can decrease EEBPR and the risk for hypertension by decreasing arterial resistance. We studied the effects of aerobic training on the submaximal exercise blood pressure (BP) of eight normotensive young adult African-American men with an EEBPR. Subjects were trained on a stationary bicycle at an intensity of 70% peak oxygen uptake (VO2peak) for 30 min, three times per week, for 8 weeks. BP, heart rate, cardiac output (CO), stroke volume (SV) and total peripheral vascular resistance (TPR) were measured at rest and during submaximal exercise at a work intensity of 50% VO2peak. Significance of the training effects were evaluated by comparing the pre- and post-training measures (t-test, p < 0.05). A 15% post-training increase in VO2peak (34.6 ± 1.4 to 40 ± 1.4 ml/kg/min) and a 9.5 ml post-training increase in mean resting stroke volume were found. A 16.2 mmHg decrement in mean systolic BP, an 11.5 mmHg decrement in mean diastolic BP, a 120 dyne/s/cm5 decrement in TPR and a 1.2 l/min increase in CO were detected during the post-training submaximal exercise tests. These results suggest that reductions in TPR may attenuate the EEBPR of normotensive African-American males following an 8-week training regimen of stationary bicycling at 70% VO2peak. Aerobic exercise training may, therefore, reduce the risk of hypertension in normotensive African-American males by the mechanism of a reduction in TPR. Because of the limited number of subjects, the results of this study should be interpreted cautiously pending confirmation by a larger controlled trial. PMID:12361191

  2. Growth characteristics of freeze-tolerant baker's yeast Saccharomyces cerevisiae AFY in aerobic batch culture.

    PubMed

    Ji, Meng; Miao, Yelian; Chen, Jie Yu; You, Yebing; Liu, Feilong; Xu, Lin

    2016-01-01

    Saccharomyces cerevisiae AFY is a novel baker's yeast strain with strong freeze-tolerance, and can be used for frozen-dough processing. The present study armed to clarify the growth characteristics of the yeast AFY. Aerobic batch culture experiments of yeast AFY were carried out using media with various initial glucose concentrations, and the culture process was analyzed kinetically. The growth of the yeast AFY exhibited a diauxic pattern with the first growth stage consuming glucose and the second growth stage consuming ethanol. The cell yield decreased with increasing initial glucose concentration in the first growth stage, and also decreased with increasing initial ethanol concentration in the second growth stage. In the initial glucose concentration range of 5.0-40.0 g/L, the simultaneous equations of Monod equation, Luedeking-Piret equation and pseudo-Luedeking-Piret equation could be used to describe the concentrations of cell, ethanol and glucose in either of the two exponential growth phases. At the initial glucose concentrations of 5.0, 10.0 and 40.0 g/L, the first exponential growth phase had a maximal specific cell growth rate of 0.52, 0.98 and 0.99 h(-1), while the second exponential growth phase had a maximal specific cell growth rate of 0.11, 0.06 and 0.07 h(-1), respectively. It was indicated that the efficiency of the yeast production could be improved by reducing the ethanol production in the first growth stage. PMID:27186467

  3. Short-term low-intensity blood flow restricted interval training improves both aerobic fitness and muscle strength.

    PubMed

    de Oliveira, M F M; Caputo, F; Corvino, R B; Denadai, B S

    2016-09-01

    The present study aimed to analyze and compare the effects of four different interval-training protocols on aerobic fitness and muscle strength. Thirty-seven subjects (23.8 ± 4 years; 171.7 ± 9.5 cm; 70 ± 11 kg) were assigned to one of four groups: low-intensity interval training with (BFR, n = 10) or without (LOW, n = 7) blood flow restriction, high-intensity interval training (HIT, n = 10), and combined HIT and BFR (BFR + HIT, n = 10, every session performed 50% as BFR and 50% as HIT). Before and after 4 weeks training (3 days a week), the maximal oxygen uptake (VO2max ), maximal power output (Pmax ), onset blood lactate accumulation (OBLA), and muscle strength were measured for all subjects. All training groups were able to improve OBLA (BFR, 16%; HIT, 25%; HIT + BFR, 22%; LOW, 6%), with no difference between groups. However, VO2max and Pmax improved only for BFR (6%, 12%), HIT (9%, 15%) and HIT + BFR (6%, 11%), with no difference between groups. Muscle strength gains were only observed after BFR training (11%). This study demonstrates the advantage of short-term low-intensity interval BFR training as the single mode of training able to simultaneously improve aerobic fitness and muscular strength. PMID:26369387

  4. Blood pressure changes following aerobic exercise in Caucasian and Chinese descendants.

    PubMed

    Sun, P; Yan, H; Ranadive, S M; Lane, A D; Kappus, R M; Bunsawat, K; Baynard, T; Li, S; Fernhall, B

    2015-03-01

    Acute aerobic exercise produces post-exercise hypotension (PEH). Chinese populations have lower prevalence of cardiovascular disease compared to Caucasians. PEH may be associated cardiovascular disease through its influence on hypertension. The purpose of this study was to compare PEH between Caucasian and Chinese subjects following acute aerobic exercise. 62 (30 Caucasian and 32 Chinese, 50% male) subjects underwent measurement of peripheral and central hemodynamics as well as arterial and cardiac evaluations, 30 min and 60 min after 45 min of treadmill exercise. Caucasians exhibited significantly higher baseline BP than the Chinese. While the reduction in brachial artery systolic BP was greater in Caucasian than in the Chinese, there was no difference in changes in carotid systolic BP between the groups. The increase in cardiac output and heart rate was greater in the Chinese than Caucasians, but total peripheral resistance and leg pulse wave velocity decreased by a similar magnitude in the Chinese and Caucasian subjects. We conclude that acute aerobic exercise produces a greater magnitude of PEH in peripheral systolic BP in Caucasian compared to Chinese subjects. The different magnitude in PEH was caused by the greater increase in cardiac output mediated by heart rate, with no change in stroke volume. It is possible that initial BP differences between races influenced the findings. PMID:25329430

  5. Bacteriological analysis of blood culture isolates from neonates in a tertiary care hospital in India.

    PubMed

    Kumhar, Ghanshyam D; Ramachandran, V G; Gupta, Piyush

    2002-12-01

    This study was undertaken to determine the profile and antibiotic sensitivity patterns of aerobic isolates from blood cultures of neonates in a tertiary care hospital in New Delhi, India. All blood culture reports (n = 1,828), obtained during August 1995-September 1996 from newborns admitted to the Department of Pediatrics and the Neonatal Intensive Care Unit at the University College of Medical Sciences and GTB Hospital, Delhi, were analyzed, and the sensitivity patterns were recorded. The positivity of blood culture was 42% (770/1,828). Most (93.2%) bactaeremic episodes were caused by a single organism, while polymicrobial aetiology was observed in 52 (6.8%) cases. Gram-negative organisms were isolated in 493 (60%) of 823 cases, with Klebsiella (33.8%), Enterobacter (7.5%), Alcaligenes faecalis (4.9%), and Escherichia coli (4.6%) being the common microbes. Staphylococcus aureus (24.4%), followed by coagulase-negative staphylococci (7.9%), were the major Gram-positive isolates. Most (80%) Gram-positive isolates were sensitive to vancomycin, and 50-75% of the Gram-negative isolates were sensitive to ciprofloxacin and amikacin. It is concluded that Klebsiella and Staphylococcus aureus remain the principal organisms responsible for neonatal sepsis in a tertiary care setting. PMID:12659415

  6. Bacteriological culture of blood from critically ill neonatal calves.

    PubMed Central

    Fecteau, G; Van Metre, D C; Paré, J; Smith, B P; Higgins, R; Holmberg, C A; Jang, S; Guterbock, W

    1997-01-01

    The objectives of this study were to estimate the prevalence of bacteremia in critically ill, neonatal calves with severe diarrhea or depression, and to describe the variety of bacteria involved. Two studies were conducted in the summers of 1991 and 1993 involving 190 neonatal calves, 1-day to 19-days-old. Bacteremia was detected by blood culture in 31% (28/90) of calves in study 1, and in 24% (19/79) of ill calves and 0% (0/21) of control calves in study 2. Bacteria cultured from blood included Escherichia coli (51% of all isolates), other gram-negative enterics (25.5%), gram-negative anaerobes (5.9%), gram-positive cocci (11.8%), and gram-positive rods (5.9%). Among clinically ill calves, the average age was significantly lower in the blood culture-negative group (5.5 d) than in the blood culture-positive group (7.5 d) (P = 0.004). Mean serum IgG concentration was significantly (P = 0.0001) lower in blood culture-positive calves (1.146 g/L) than in blood culture-negative calves (3.077 g/L). The mortality rate was significantly (P < 0.0001) higher in the blood culture-positive group (57.4%) than in the blood culture-negative group (15.1%). Bacteremia appeared to be a frequent entity in this particular rearing situation. Early recognition of the problem, as well as appropriate treatment, may be beneficial in increasing survival rates. Results also support the need to address the failure of passive transfer of maternal antibodies to prevent bacteremia in calves. Images Figure 1. PMID:9028592

  7. Combined effects of aerobic exercise and l-arginine ingestion on blood pressure in normotensive postmenopausal women: A crossover study.

    PubMed

    Puga, Guilherme M; de P Novais, Iane; Katsanos, Christos S; Zanesco, Angelina

    2016-04-15

    After menopause the incidence of cardiovascular diseases increases in women. A decrease in nitric oxide (NO) bioavailability has been pointed out to play a major role in this phenomenon. Since it is believed that l-arginine administration could improve NO bioavailability, the aim of this study was to examine the effects of acute l-arginine administration associated with aerobic exercise on blood pressure (BP), redox state and inflammatory biomarkers in normotensive postmenopausal women (NPW). Sixteen volunteers (57±6yr) were subjected to four experimental sessions (crossover design): arginine+exercise (A-E); arginine (ARG); exercise+placebo (EXE); control (CON). Each session was initiated with either 9g of l-arginine ingestion (ARG or A-E days), placebo (EXE day), or nothing (CON day). The participants performed 30min of aerobic exercise (A-E and EXE days) or sitting rest (CON and ARG days). Blood samples were collected before each session and 45min after the intervention. Office BP and ambulatory blood pressure monitoring (ABPM) were evaluated. NO/cGMP pathway, redox state and inflammatory biomarkers were measured. Systolic BP decreased during the 24-hour in A-E and EXE sessions. However, diastolic BP reduced only in A-E session. No changes were found in the biomarkers concentrations. In conclusion, the association was effective in lowering diastolic BP in NPW. Additionally, physical exercise alone promoted a long lasting effect on systolic BP measured by ABPM in this population, although this beneficial effect was not associated with changes in the cardio-inflammatory biomarkers. Possibly, other factors such as neural influences could be mediating this effect. PMID:26972606

  8. Transport and metabolism of fumaric acid in Saccharomyces cerevisiae in aerobic glucose-limited chemostat culture.

    PubMed

    Shah, Mihir V; van Mastrigt, Oscar; Heijnen, Joseph J; van Gulik, Walter M

    2016-04-01

    Currently, research is being focused on the industrial-scale production of fumaric acid and other relevant organic acids from renewable feedstocks via fermentation, preferably at low pH for better product recovery. However, at low pH a large fraction of the extracellular acid is present in the undissociated form, which is lipophilic and can diffuse into the cell. There have been no studies done on the impact of high extracellular concentrations of fumaric acid under aerobic conditions in S. cerevisiae, which is a relevant issue to study for industrial-scale production. In this work we studied the uptake and metabolism of fumaric acid in S. cerevisiae in glucose-limited chemostat cultures at a cultivation pH of 3.0 (pH < pK). Steady states were achieved with different extracellular levels of fumaric acid, obtained by adding different amounts of fumaric acid to the feed medium. The experiments were carried out with the wild-type S. cerevisiae CEN.PK 113-7D and an engineered S. cerevisiae ADIS 244 expressing a heterologous dicarboxylic acid transporter (DCT-02) from Aspergillus niger, to examine whether it would be capable of exporting fumaric acid. We observed that fumaric acid entered the cells most likely via passive diffusion of the undissociated form. Approximately two-thirds of the fumaric acid in the feed was metabolized together with glucose. From metabolic flux analysis, an increased ATP dissipation was observed only at high intracellular concentrations of fumarate, possibly due to the export of fumarate via an ABC transporter. The implications of our results for the industrial-scale production of fumaric acid are discussed. Copyright © 2015 John Wiley & Sons, Ltd. PMID:26683700

  9. Rapid identification of Candida species in blood cultures by a clinically useful PCR method.

    PubMed Central

    Shin, J H; Nolte, F S; Morrison, C J

    1997-01-01

    Widespread use of fluconazole for the prophylaxis and treatment of candidiasis has led to a reduction in the number of cases of candidemia caused by Candida albicans but has also resulted in the emergence of candidemias caused by innately fluconazole-resistant, non-C. albicans Candida species. Given the fulminant and rapidly fatal outcome of acute disseminated candidiasis, rapid identification of newly emerging Candida species in blood culture is critical for the implementation of appropriately targeted antifungal drug therapy. Therefore, we used a PCR-based assay to rapidly identify Candida species from positive blood culture bottles. This assay used fungus-specific, universal primers for DNA amplification and species-specific probes to identify C. albicans, C. krusei, C. parapsilosis, C. tropicalis, or C. glabrata amplicons. It also used a simpler and more rapid (1.5-h) sample preparation technique than those described previously and used detergent, heat, and mechanical breakage to recover Candida species DNA from blood cultures. A simple and rapid (3.5-h) enzyme immunosorbent assay (EIA)-based format was then used for amplicon detection. One hundred fifty blood culture bottles, including 73 positive blood culture bottle sets (aerobic and anaerobic) from 31 patients with candidemia, were tested. The combined PCR and EIA methods (PCR-EIA) correctly identified all Candida species in 73 blood culture bottle sets, including bottles containing bacteria coisolated with yeasts and 3 cultures of samples from patients with mixed candidemias originally identified as single-species infections by routine phenotypic identification methods. Species identification time was reduced from a mean of 3.5 days by routine phenotypic methods to 7 h by the PCR-EIA method. No false-positive results were obtained for patients with bacteremias (n = 18), artificially produced non-Candida fungemias (n = 3), or bottles with no growth (n = 20). Analytical sensitivity was 1 cell per 2-microl

  10. Culturable Aerobic and Facultative Anaerobic Intestinal Bacterial Flora of Black Cobra (Naja naja karachiensis) in Southern Pakistan

    PubMed Central

    Iqbal, Junaid; Sagheer, Mehwish; Tabassum, Nazneen; Siddiqui, Ruqaiyyah; Khan, Naveed Ahmed

    2014-01-01

    Using morphological analysis and biochemical testing, here for the first time, we determined the culturable gut bacterial flora (aerobes and facultative anaerobes) in the venomous Black Cobra (Naja naja karachiensis) from South Asia. The findings revealed that these snakes inhabit potentially pathogenic bacteria including Serratia marcescens, Pseudomonas aeruginosa, Shewanella putrefaciens, Aeromonas hydrophila, Salmonella sp., Moraxella sp., Bacillus sp., Ochrobactrum anthropi, and Providencia rettgeri. These findings are of concern, as injury from snake bite can result in wound infections and tissue necrosis leading to sepsis/necrotizing fasciitis and/or expose consumers of snake meat/medicine in the community to infections. PMID:25002979

  11. Culturable Aerobic and Facultative Anaerobic Intestinal Bacterial Flora of Black Cobra (Naja naja karachiensis) in Southern Pakistan.

    PubMed

    Iqbal, Junaid; Sagheer, Mehwish; Tabassum, Nazneen; Siddiqui, Ruqaiyyah; Khan, Naveed Ahmed

    2014-01-01

    Using morphological analysis and biochemical testing, here for the first time, we determined the culturable gut bacterial flora (aerobes and facultative anaerobes) in the venomous Black Cobra (Naja naja karachiensis) from South Asia. The findings revealed that these snakes inhabit potentially pathogenic bacteria including Serratia marcescens, Pseudomonas aeruginosa, Shewanella putrefaciens, Aeromonas hydrophila, Salmonella sp., Moraxella sp., Bacillus sp., Ochrobactrum anthropi, and Providencia rettgeri. These findings are of concern, as injury from snake bite can result in wound infections and tissue necrosis leading to sepsis/necrotizing fasciitis and/or expose consumers of snake meat/medicine in the community to infections. PMID:25002979

  12. Comparison of fumerate-pyruvate media and beef extract media for aerobically culturing Campylobacter species

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Media supplemented with fumarate, pyruvate, and a vitamin-mineral solution or with beef extract were compared for the ability to support aerobic growth of Campylobacter. Basal broth composed of tryptose, yeast extract, bicarbonate, and agar was supplemented with 30 mM fumarate, 100 mM pyruvate, and ...

  13. Oxygen uptake, heart rate and blood lactate concentration during a normal training session of an aerobic dance class.

    PubMed

    De Angelis, M; Vinciguerra, G; Gasbarri, A; Pacitti, C

    1998-07-01

    The aim of this research was to investigate the physiological responses and, in particular, the participation of lactic acid anaerobic metabolism in aerobic dance, which is claimed to be pure aerobic exercise. In contrast to previous studies, that have put subjects in very unfamiliar situations, the parameters were monitored in the familiar context of gymnasium, practice routine and habitual instructor. A group of 30 skilled fairly well-trained women performed their usual routine, a combination of the two styles: low (LI) and high impact (HI), and were continuously monitored for heart rate (HR) and every 8 min for blood lactate concentration ([La-]b). Of the group, 15 were tested to determine their maximal aerobic power (VO2max) using a cycle-ergometer. They were also monitored during the routine for oxygen uptake (VO2) by a light telemetric apparatus. The oxygen pulses of the routine and of the corresponding exercise intensity in the incremental test were not statistically different. The mean values in the exercise session were: peak HR 92.8 (SD 7.8)% of the subject's maximal theoretical value, peak VO2 99.5 (SD 12.4)% of VO2max, maximal [La-]b 6.1 (SD 1.7) mmol x l(-1), and mean 4.8 (SD 1.3) mmol x l(-1). Repeated measures ANOVA found statistically significant differences between the increasing [La-]b values (P < 0.001). In particular, the difference between the [La-]b values at the end of the mainly LI phase and those of the LI-HI combination phase, and the difference between the samples during the combination LI-HI phase were both statistically significant (both P = 0.002 and P = 0.002). The similar oxygen pulses confirmed the validity of the present experiment design and the reliability of HR monitoring in this activity. The HR, VO2 and, above all, the increase of [La-]b to quite high values, showing a non steady state, demonstrated the high metabolic demand made by this activity that involved lactic acid metabolism at a much higher level than expected. PMID

  14. Culture of Piscirickettsia salmonis on enriched blood agar.

    PubMed

    Mauel, Michael J; Ware, Cynthia; Smith, Pedro A

    2008-03-01

    Piscirickettsia salmonis is the etiologic agent of piscirickettsiosis, an economically significant disease of fish. Isolation of P. salmonis by culturing on fish cell lines has been the standard technique since the initial isolation of the organism. The ability to grow P. salmonis on artificial media would relieve facilities of the cost of maintaining cell lines, permit isolation at fish culture sites with fewer contamination problems, and allow easier transport of isolates to diagnostic facilities for confirmation assays. This report describes the successful culture of P. salmonis on enriched blood agar. PMID:18319435

  15. Effect of chlorate, molybdate, and shikimic acid on Salmonella Typhimurium in aerobic and anaerobic cultures

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two studies were conducted to examine the effects of shikimic acid (60 µg/mL) and(or) molybdate (1 mM) on the sensitivity of Salmonella enterica serovar Typhimurium to sodium chlorate (5 mM) during anaerobic (90% N2:5% CO2:5% H2) or aerobic growth in brain heart infusion broth supplemented with 5 mM...

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

    SciTech Connect

    Nogueira, N.; Chaplan, S.; Tydings, J.D.; Unkeless, J.; Cohn, Z.

    1981-03-01

    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 (1).

  17. Comparison of aerobic denitrifying activity among three cultural species with various carbon sources.

    PubMed

    Otani, Y; Hasegawa, K; Hanaki, K

    2004-01-01

    Abilities of three aerobic denitrifiers such as Alcaligenes faecalis, Microvirgula aerodenitrificans and Paracoccus pantotrophus were compared from the viewpoints of nitrate removal efficiency and organic matter utilization. First, the effect of carbon source was investigated. Although nitrate reduction was observed in all strains under aerobic conditions, a change of carbon source considerably affected the denitrification ability. In the case of P. pantotrophus, nitrate and nitrite were completely removed in three days under sodium acetate or leucine as a carbon source. In the case of A. faecalis, sufficient nitrate removal was observed only when sodium acetate or ethanol was added. P. pantotrophus and A. faecalis showed a higher ability of nitrate removal than that of M. aerodenitrificans. Therefore, P. pantotrophus was selected in order to investigate the effects of concentration and repetitive addition of carbon. Sodium acetate was used as a sole carbon source. Nitrate was not reduced when the carbon concentration was below 500 mgC/L. However, when carbon source was added repeatedly, nitrate was reduced under 100 mgC/L after the optical density of the bacterium reached above 1.0. This result indicated that a high enough level of bacterial density was necessary to express aerobic denitrification activity. PMID:15566182

  18. Effects of aerobic exercise on blood glucose in continuous ambulatory peritoneal dialysis patients

    PubMed Central

    Shahgholian, Nahid; KarimiFard, Ozra; Shahidi, Shahrzad

    2015-01-01

    Background: Peritoneal dialysis has a number of complications including increased blood glucose. Although exercise has been suggested to resolve this complication, most patients are not active. The present study aimed at determining the effects of twice-weekly, 40-min sessions of pedaling on a stationary bicycle on mean fasting blood sugar (FBS) and 2-h postprandial blood sugar (PPBS) among continuous ambulatory peritoneal dialysis patients. Materials and Methods: In this clinical trial, convenience sampling was used to select 22 patients [age: 51.4 (12.3) years] undergoing continuous ambulatory peritoneal dialysis [mean duration: 12.5 (8.5) months] from university hospitals in Isfahan, Iran. The subjects were randomly divided into two groups (test and control). The test group participated in an 8-week exercise program in which they pedaled a stationary bicycle with an intensity of four on Borg Scale of Perceived Exertion. FBS and PPBS were measured at baseline and at the end of the 8th and 16th sessions of exercise. Data were analyzed with Student's t-test and repeated measures analysis of variance. Results: After the eighth session, the mean FBS and PPBS levels were lower in the test group than in the control group. However, the differences were not statistically significant. After 16 sessions of exercise, the mean FBS and PPBS levels in the intervention group were significantly less than the in control group. Conclusions: Forty minutes of pedaling on a stationary bicycle for two times a week can significantly reduce mean FBS and PPBS levels in continuous ambulatory peritoneal dialysis patients. PMID:25878690

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

    SciTech Connect

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

    1989-03-01

    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.

  20. Can a Single Session of a Community-Based Group Exercise Program Combining Step Aerobics and Bodyweight Resistance Exercise Acutely Reduce Blood Pressure?

    PubMed Central

    Mendes, Romeu; Sousa, Nelson; Garrido, Nuno; Cavaco, Braulio; Quaresma, Luís; Reis, Victor Machado

    2014-01-01

    This study aimed to analyze the acute effects of a single session of a community-based group exercise program combining step aerobics and bodyweight resistance exercise on blood pressure in healthy young adult women. Twenty-three healthy young adult women (aged 31.57 ± 7.87 years) participated in two experimental sessions (exercise and control) in a crossover study design. Blood pressure was monitored before, immediately after and at 10, 20 and 30 min of recovery. The exercise session consisted of four phases: 1) a warm-up (5 min of dance aerobics); 2) aerobic exercise training (30 min of step aerobics); 3) resistance exercise training (six sets of 12 repetitions of three bodyweight exercises in a circuit mode, 10 min); and 4) a cool-down (5 min of breathing and flexibility exercises); totaling 50 min of duration. Systolic blood pressure after exercise was significantly lower compared to control at the 10th min (−10.83 ± 2.13 vs. −2.6 ± 2.13 mmHg; p = 0.009), 20th min (−11.26 ± 2.13 vs. −3.04 ± 2.13 mmHg; p = 0.009) and 30th min of recovery (−10.87 ± 2.39 vs. −0.48 ± 2.39 mmHg; p = 0.004). A single session of a community-based group exercise program combining step aerobics and bodyweight resistance exercise was effective in inducing significant post-exercise hypotension in healthy young adult women. This type of low-cost exercise interventions may have an important role in the prevention of cardiovascular diseases and in community health promotion. PMID:25713644

  1. [Research advances in aerobic denitrifiers].

    PubMed

    Wang, Wei; Cai, Zu-cong; Zhong, Wen-hui; Wang, Guo-xiang

    2007-11-01

    This paper reviewed the varieties and characteristics of aerobic denitrifiers, their action mechanisms, and the factors affecting aerobic denitrification. Aerobic denitrifiers mainly include Pseudomonas, Alcaligenes, Paracoccus and Bacillus, which are either aerobic or facultative aerobic, and heterotrophic. They can denitrify under aerobic conditions, with the main product being N2O. They can also convert NH4+ -N to gas product. The nitrate reductase which catalyzes the denitrification is periplasmic nitrate reductase rather than membrane-bound nitrate reductase. Dissolved oxygen concentration and C/N ratio are the main factors affecting aerobic denitrification. The main methods for screening aerobic denitrifiers, such as intermittent aeration and selected culture, were also introduced. The research advances in the application of aerobic denitrifiers in aquaculture, waste water processing, and bio-degradation of organic pollutants, as well as the contributions of aerobic denitrifiers to soil nitrogen emission were summarized. PMID:18260473

  2. Orange juice improved lipid profile and blood lactate of overweight middle-aged women subjected to aerobic training

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study evaluated the influence of regular consumption of orange juice associated with aerobic exercise on the lipid profile of middle aged women, previously sedentary. Twenty-six women, 30 to 55 years old, volunteered to consume orange juice daily for 3 months and participate in an aerobic train...

  3. [Microbial maps and blood cultures in acute leukemia].

    PubMed

    Rossi, M; Roberti, M G; Paolino, F

    1976-12-29

    Microbial maps were performed taking swabs from nose, pharinx, external auditory meatus, groin, vagina, sputum and urine cultures in 69 cases of acute leukaemia, in order: to assess the germs' incidence in an "open ward" department; to eliminate the most dangerous pathogens with local treatment or with a selective therapy without broad-specturm antibiotics; to check, in the 43 cases followed from onset, the changes occurring during the admission and the disease progression; to collect data for comparison with a "sterile" ward. The local decontamination had only a temporary effect. During the course of the disease new, particularly dangerous, pathogens were cultured. Blood cultures were positive in 15% of the patients with fever at the onset of the disease, and in 36.9% of the patients with fever during the disease progression. These values were virtually the same as those observed in the acute stage of C.M.L. (35.7%). In akute leukaemia E. coli (35%) was the most common, followed by P. aeruginosa (20%), Klebsiella (15%), S. alpha haemolyticus (10%) and others. There was little or no relationship between the germs in the maps and those in the blood cultures, though it must be remembered that no stool cultures were examined. PMID:1035410

  4. First Report of Clostridium lavalense Isolated in Human Blood Cultures

    PubMed Central

    Bourque, Christine; Thibault, Louise; Côté, Jean-Charles; Domingo, Marc-Christian

    2016-01-01

    An 88-year-old man was admitted to the hospital with worsening malaise, fever, and weakness. Anaerobic blood culture bottles revealed the presence of an anaerobic, Gram-positive sporulated bacillus. Empirical antibiotherapy with intravenous piperacillin-tazobactam was initiated. The patient defervesced after four days and was switched to oral amoxicillin on his 6th day of antibiotic therapy and later discharged from the hospital. Four months later, he had recovered. The bacterium was initially identified as Clostridium butyricum using anaerobic manual identification panel. 16S rRNA gene sequence and phylogenetic analysis showed the bacterium to be Clostridium lavalense, a recently described species with no previously published case of isolation in human diagnostic samples so far. This is the first report of Clostridium lavalense isolation from human blood cultures. Further studies are needed in order to elucidate the role of Clostridium lavalense in human disease and its virulence factors. PMID:27478446

  5. Rapid Detection of ESBL-Producing Enterobacteriaceae in Blood Cultures

    PubMed Central

    Dortet, Laurent; Poirel, Laurent

    2015-01-01

    We rapidly identified extended-spectrum β-lactamase (ESBL) producers prospectively among 245 gram-negative bacilli–positive cultured blood specimens using the Rapid ESBL Nordmann/Dortet/Poirel test and direct bacterial identification using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. This combination identified ESBL-producing Enterobacteriaceae within 30 min and had high predictive values. PMID:25695535

  6. Rapid Identification of Bacteria from Positive Blood Culture Bottles by Use of Matrix-Assisted Laser Desorption-Ionization Time of Flight Mass Spectrometry Fingerprinting▿ †

    PubMed Central

    Christner, Martin; Rohde, Holger; Wolters, Manuel; Sobottka, Ingo; Wegscheider, Karl; Aepfelbacher, Martin

    2010-01-01

    Early and adequate antimicrobial therapy has been shown to improve the clinical outcome in bloodstream infections (BSI). To provide rapid pathogen identification for targeted treatment, we applied matrix-assisted laser desorption-ionization time of flight (MALDI-TOF) mass spectrometry fingerprinting to bacteria directly recovered from blood culture bottles. A total of 304 aerobic and anaerobic blood cultures, reported positive by a Bactec 9240 system, were subjected in parallel to differential centrifugation with subsequent mass spectrometry fingerprinting and reference identification using established microbiological methods. A representative spectrum of bloodstream pathogens was recovered from 277 samples that grew a single bacterial isolate. Species identification by direct mass spectrometry fingerprinting matched reference identification in 95% of these samples and worked equally well for aerobic and anaerobic culture bottles. Application of commonly used score cutoffs to classify the fingerprinting results led to an identification rate of 87%. Mismatching mostly resulted from insufficient bacterial numbers and preferentially occurred with Gram-positive samples. The respective spectra showed low concordance to database references and were effectively rejected by score thresholds. Spiking experiments and examination of the respective study samples even suggested applicability of the method to mixed cultures. With turnaround times around 100 min, the approach allowed for reliable pathogen identification at the day of blood culture positivity, providing treatment-relevant information within the critical phase of septic illness. PMID:20237093

  7. Doing it right the first time: quality improvement and the contaminant blood culture.

    PubMed Central

    Weinbaum, F I; Lavie, S; Danek, M; Sixsmith, D; Heinrich, G F; Mills, S S

    1997-01-01

    The aim of the project was to determine whether the rate of contaminant blood cultures could be reduced by using a team of dedicated phlebotomists. Comparisons were made between adult patients requiring blood cultures for suspected bacteremia on medical and surgical units before and after the introduction and withdrawal of a dedicated blood culture team. The results showed that a significant reduction in the contaminant blood culture rate was achieved by the blood culture team (P < 0.001; chi(2) test). Therefore, in our experience, the rate of contaminant blood cultures can be reduced in a teaching hospital by using a team of dedicated phlebotomists. Calculations made with our data and those published by others suggest that cost savings from reducing false-positive blood cultures are greater than the cost of the blood culture team. PMID:9041389

  8. Bacteremia during dacryocystorhinostomy: results of intra-operative blood cultures

    PubMed Central

    2014-01-01

    Background The aims of the study are to assess the prevalence of bacteremia during dacryocystorhinostomy (DCR) and to assess whether there is a need for post-operative prophylaxis. Prospective interventional study of 52 consecutive dacryocystorhinostomy performed in 50 patients over a period of 1 year from 2013 to 2014. Blood was drawn under strict aseptic conditions during two separate time points: fashioning of the nasal mucosal and creation of lacrimal sac flaps. The blood was inoculated into two blood culture bottles: the dual media as well as Columbia broth. Following withdrawal of blood, all patients received an intraoperative single dose of a cephalosporin antibiotic. Clean cases of primary acquired nasolacrimal duct obstructions (PANDO) without any sac discharge upon marsupialization (22%, 11/50) were not prescribed routine post-operative prophylaxis, whereas the remaining were prescribed oral antibiotics for 5 days. Results The mean age of patients was 41 years (range, 4–61 years). The most common diagnosis (70%, 35/50) was primary acquired nasolacrimal duct obstruction. Acute dacryocystitis was noted in 12% (6/50). External DCR was performed in 65% (34/52) and endoscopic DCR in 35% (18/52) of the cases. All the blood cultures were uniformly negative both in terms of abnormal physical changes in media as well subcultures; 22% (11/50) did not receive post-operative antibiotic prophylaxis. None of the patients developed any signs of wound infections. The anatomical and functional success rate was achieved in 98%. Conclusions This study did not find any intraoperative bacteremia during dacryocystorhinostomy and that none had wound infection irrespective of post-operative prophylaxis. PMID:25320650

  9. Ability of Cecal Cultures to Inhibit Growth of Salmonella Typhimurium during Aerobic Incubation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction: Poultry can serve as reservoirs for Salmonella; however, chicks provided cultures of cecal bacteria develop resistance to colonization by Salmonella. Research has indicated that cecal bacteria metabolize organic acids to produce substances that inhibit Salmonella growth. Purpose: The...

  10. Enrichment culture for the isolation of Campylobacter spp: Effects of incubation conditions and the inclusion of blood in selective broths.

    PubMed

    Williams, Lisa K; Jørgensen, Frieda; Grogono-Thomas, Rose; Humphrey, Tom J

    2009-03-31

    Isolation of Campylobacter spp. using enrichment culture is time consuming and complex. Reducing the time taken to confirm the presence or absence of Campylobacter spp. would have many advantages for diagnostic, commercial and research applications. Rapid techniques such as real-time PCR can detect campylobacters from complex samples but blood in enrichment culture can inhibit the PCR reaction, if applied directly to enriched samples. The aim of this study was to investigate the effect of blood in enrichment culture on the isolation of campylobacters from chicken caeca, carcass rinses and bootsock (gauze sock walked through a broiler chicken house) samples using Bolton broth. The effect of incubation temperature (37 degrees C or 41.5 degrees C for 48 h, or 37 degrees C for 4 h then transfer to 41.5 degrees C for 44 h) and method of generating atmosphere (incubation of container in jar gassed with microaerobic atmosphere or incubation of container with small headspace and tightly screwed lid in an aerobic atmosphere) with and without blood on isolation from chicken carcass rinses and chicken faeces was also investigated. The presence of blood in enrichment culture did not improve the isolation of campylobacters from chicken faeces or bootsock samples but significantly improved recovery from chicken carcass rinse samples. There was no significant effect of the method used to generate incubation atmosphere. Isolation rates did also not depend significantly on whether broths were incubated at 37 or 41.5 degrees C for 24 or 48 h. Overall, the presence of blood in such media is not essential, although isolation can vary depending on sample type and enrichment method used. PMID:19217181

  11. Effects of aerobic dance training on blood pressure in individuals with uncontrolled hypertension on two antihypertensive drugs: a randomized clinical trial.

    PubMed

    Maruf, Fatai Adesina; Akinpelu, Aderonke Omobonike; Salako, Babatunde Lawal; Akinyemi, Joshua Odunayo

    2016-04-01

    There is a dearth of reports on possible additive blood pressure (BP)-reducing effect of aerobic exercise on antihypertensive drug in humans. This study investigated the additive BP-reducing effect of aerobic exercise on BP in individuals with uncontrolled hypertension. In this 12-week double-blind study, 120 new-diagnosed individuals with mild-to-moderate hypertension were randomized to receive coamilozide + 5/10 mg of amlodipine + aerobic dance or coamilozide + 5/10 mg of amlodipine alone. Forty-five and 43 participants in exercise and control groups, respectively, completed the 12-week intervention. Addition of aerobic exercise to antihypertensive drug therapy significantly reduced systolic BP (7.1 mm Hg [95% confidence interval: 5.0, 9.3]; P < .001) and diastolic BP (1.7 mm Hg [95% confidence interval: 0.4, 3.0]; P = .009) at 12 weeks. BP control rate differed significantly between exercise (53.9%) and control (35.3%) groups, P < .001. Postintervention, proportion of participants in exercise group who had their number of antihypertensive drug reduced to one (20.3%) differed from that in control group (11.1%); (χ(2) = 11.0; P = .001). Combination of aerobic dance and antihypertensive drugs reduces number of antihypertensive drugs needed to achieve BP control and enhances BP control in individuals with hypertension on two antihypertensive drugs. PMID:26948962

  12. Evaluation of a Novel Dry Sheet Culture Method for Rapid Enumeration of Total Aerobic Count in Foods.

    PubMed

    Teramura, Hajime; Iwasaki, Mihoko; Ushiyama, Masashi; Ogihara, Hirokazu

    2015-10-01

    A novel dry sheet culture method (Sanita-kun ACplus; SkACp) for rapid enumeration of total viable count has been developed. This rehydrated plate system comprises an adhesive sheet, nonwoven fabric coated with nutrients, and two types of water absorption polymers. In addition, SkACp facilitates methods for both rapid count (rapid mode: 24-h incubation) and accurate enumeration (standard mode: 48-h incubation) because it not only contains conventional 2,3,5-triphenyltetrazolium chloride but also contains two kinds of new tetrazolium salts for rapid and accurate enumeration of total aerobic count. When SkACp was assessed with 91 microorganisms, 87 strains (95.6%), excluding lactic acid and psychrotrophic bacteria, formed red-colored colonies within 24 h, whereas all microorganisms tested formed colonies within 48 h. The SkACp method, with both 24 and 48 h of incubation, was compared with plate count agar (PCA) and 3M Petrifilm AC (PAC) by using 107 naturally contaminated foods. For all foods tested (n = 107), the linear correlation coefficients of 48-h counts on SkACp compared with PCA and PAC were 0.98 and 0.75, respectively, while the 24-h counts on SkACp compared with PCA and PAC were 0.77 and 0.96, respectively. For foods tested, excluding yogurt and lactic beverages ( n = 101), the linear correlation coefficients of 48-h counts on SkACp compared with PCA and PAC were 0.98 and 0.96, respectively, while the 24-h counts on SkACp compared with PCA and PAC were 0.96 and 0.95, respectively. These results demonstrated that SkACp (48 h) is a useful alternative for the enumeration of the total aerobic count for all foods, whereas SkACp (24 h) was also an effective method for rapid enumeration in foods, excluding yogurt and lactic beverages. PMID:26408139

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

    PubMed Central

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

    2016-01-01

    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

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

    PubMed

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

    2016-01-01

    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

  15. Comparative Effects of Vigorous-Intensity and Low-Intensity Blood Flow Restricted Cycle Training and Detraining on Muscle Mass, Strength, and Aerobic Capacity.

    PubMed

    Kim, Daeyeol; Singh, Harshvardhan; Loenneke, Jeremy P; Thiebaud, Robert S; Fahs, Christopher A; Rossow, Lindy M; Young, Kaelin; Seo, Dong-Il; Bemben, Debra A; Bemben, Michael G

    2016-05-01

    Kim, D, Singh, H, Loenneke, JP, Thiebaud, RS, Fahs, CA, Rossow, LM, Young, K, Seo, D-i, Bemben, DA, and Bemben, MG. Comparative effects of vigorous-intensity and low-intensity blood flow restricted cycle training and detraining on muscle mass, strength, and aerobic capacity. J Strength Cond Res 30(5): 1453-1461, 2016-Traditional high-intensity aerobic training has been shown to improve muscle protein synthesis and aerobic capacity; however, recent research indicates that low-intensity aerobic training with blood flow restriction (BFR) may have similar effects. The purpose of this study was to compare the effects of vigorous-intensity (VI) cycling vs. low-intensity cycling with BFR (LI-BFR) on muscle mass, strength, and aerobic capacity after training and subsequent detraining. Thirty-one physically active subjects were assigned to one of 3 groups: VI (n = 10, 60-70% heart rate reserve [HRR]), LI-BFR (n = 11, 30% HRR with BFR at 160-180 mm Hg), and no exercise control (n = 10, no exercise). Subjects in VI and LI-BFR cycled 3 times per week for 6 weeks (total 18 sessions). Body composition, muscle mass, strength, and aerobic capacity were measured pre, post, and after 3 weeks of detraining. A group × time interaction (p = 0.019) effect for both knee flexion and leg lean mass was found. For both VI and LI-BFR groups, knee flexion strength was significantly increased between pre and post (p = 0.024, p = 0.01) and between pre and 3 week-post (p = 0.039, p = 0.003), respectively. For the LI-BFR group, leg lean mass was significantly increased between pre and 3 week-post (p = 0.024) and between post and 3 week-post (p = 0.013). However, there were no significant differences between groups for any variables. The LI-BFR elicits an increase in the knee flexion muscle strength over time similar to the VI. An increase in the leg lean mass over time was seen in the LI-BFR, but not in VI and CON. PMID:26439780

  16. The effects of altitude/hypoxic training on oxygen delivery capacity of the blood and aerobic exercise capacity in elite athletes – a meta-analysis

    PubMed Central

    Park, Hun-young; Hwang, Hyejung; Park, Jonghoon; Lee, Seongno; Lim, Kiwon

    2016-01-01

    [Purpose] This study was designed as a meta-analysis of randomized controlled trials comparing effectiveness of altitude/hypoxic training (experimental) versus sea-level training (control) on oxygen delivery capacity of the blood and aerobic exercise capacity of elite athletes in Korea. [Methods] Databases (Research Information Service System, Korean studies Information Service System, National Assembly Library) were for randomized controlled trials comparing altitude/hypoxic training versus sea-level training in elite athletes. Studies published in Korea up to December 2015 were eligible for inclusion. Oxygen delivery capacity of the blood was quantified by red blood cell (RBC), hemoglobin (Hb), hematocrit (Hct), erythropoietin (EPO); and aerobic exercise capacity was quantified by maximal oxygen consumption (VO2max). RBC, Hb, Hct, VO2max represented heterogeneity and compared post-intervention between altitude/hypoxic training and sea-level training in elite athletes by a random effect model meta-analysis. EPO represented homogeneity and meta-analysis performed by a fixed effect model. Eight independent studies with 156 elite athletes (experimental: n = 82, control: n = 74) were included in the metaanalysis. [Results] RBC (4.499×105 cell/ul, 95 % CI: 2.469 to 6.529), Hb (5.447 g/dl, 95 % CI: 3.028 to 7.866), Hct (3.639 %, 95 % CI: 1.687 to 5.591), EPO (0.711 mU/mL, 95% CI: 0.282 to 1.140), VO2max (1.637 ml/kg/min, 95% CI: 0.599 to 1.400) showed significantly greater increase following altitude/hypoxic training, as compared with sea-level training. [Conclusion] For elite athletes in Korea, altitude/ hypoxic training appears more effective than sea-level training for improvement of oxygen delivery capacity of the blood and aerobic exercise capacity. PMID:27298808

  17. Neutralization of Antimicrobial Substances in New BacT/Alert FA and FN Plus Blood Culture Bottles

    PubMed Central

    Barousch, Wolfgang; Nehr, Marion; Kundi, Michael; Zeitlinger, Markus; Makristathis, Athanasios; Hirschl, Alexander M.

    2013-01-01

    Time to detection (TTD) in automated blood culture systems is delayed for sensitive microorganisms in the presence of antimicrobial substances and has been associated with worse outcomes for sepsis patients on inadequate empirical therapy. While resin addition removes antimicrobial substances to various degrees from blood culture media, media formulations and the blend of resins may influence performance. The BacT/Alert 3D system (bioMérieux) was investigated using the new resin-containing medium types FA Plus (aerobic) and FN Plus (anaerobic). TTD was compared between control and test bottles containing relevant bacteria or Candida albicans, with and without defined concentrations of antimicrobials. Failure of neutralization was defined as a negative blood culture on day 3. In general, growth delay was nonlinear, concentration dependent, bottle type specific, and reciprocally associated with MICs. Substance-specific serum drug concentrations corresponding to a predefined, clinically relevant 3-h delay of TTD were calculated. Where appropriate, a time interval allowing for drug elimination below this critical level was obtained by pharmacokinetic modeling. Clarithromycin, clindamycin, gentamicin, linezolid, tigecycline, vancomycin, and fluconazole were neutralized. For ciprofloxacin and piperacillin-tazobactam, which were only incompletely neutralized in combination with the most sensitive test strains, a maximum waiting time for blood draw of 1 h was determined based on pharmacokinetics. One or more test strains did not grow in bottles containing either amoxicillin-clavulanate, cefepime, cefotaxime, meropenem, or metronidazole, and we thus recommend particular caution in timing of blood draws if patients have been pretreated with these agents. PMID:23486710

  18. Cross-cultural variation in blood pressure: a quantitative analysis of the relationships of blood pressure to cultural characteristics, salt consumption and body weight.

    PubMed

    Waldron, I; Nowotarski, M; Freimer, M; Henry, J P; Post, N; Witten, C

    1982-01-01

    This study has analyzed the relationships of cross-cultural variation in blood pressure to cultural characteristics, salt consumption and body weight. The data used were blood pressures for adults in 84 groups, ratings of cultural characteristics (based on anthropological data and made by raters who had no knowledge of the blood pressure data) and, where available, salt consumption and body mass index (weight/height2). Blood pressures were higher and the slopes of blood pressure with age were greater in groups which had greater involvement in a money economy, more economic competition, more contact with people of different culture or beliefs, and more unfulfilled aspirations for a return to traditional beliefs and values. Blood pressures were also higher in groups for which the predominant family type was a nuclear or father-absent family, as opposed to an extended family. For Negroes, groups who were descended from slaves had higher blood pressures than other groups. The correlations between blood pressures and involvement in a money economy were substantial and significant even after controlling for level of salt consumption and, for men, also after controlling for body mass index. For men there were also significant partial correlations between blood pressure and salt consumption, controlling for type of economy. For women there were significant partial correlations between blood pressure and body mass index, controlling for type of economy. In conclusion, cross-cultural variation in blood pressure appears to be due to multiple factors. One contributory factor appears to be psychosocial stress due to cultural disruption, including the disruption of cooperative relationships and traditional cultural patterns which frequently occurs during economic modernization. In addition, both the protective effects of very low salt consumption in some groups and differences in body weight appear to contribute to cross-cultural variation in blood pressure. PMID:7079796

  19. What proportion of Salmonella Typhi cases are detected by blood culture? A systematic literature review.

    PubMed

    Mogasale, Vittal; Ramani, Enusa; Mogasale, Vijayalaxmi V; Park, JuYeon

    2016-01-01

    Blood culture is often used in definitive diagnosis of typhoid fever while, bone marrow culture has a greater sensitivity and considered reference standard. The sensitivity of blood culture measured against bone marrow culture results in measurement bias because both tests are not fully sensitive. Here we propose a combination of the two cultures as a reference to define true positive S. Typhi cases. Based on a systematic literature review, we identified ten papers that had performed blood and bone marrow culture for S. Typhi in same subjects. We estimated the weighted mean of proportion of cases detected by culture measured against true S. Typhi positive cases using a random effects model. Of 529 true positive S. Typhi cases, 61 % (95 % CI 52-70 %) and 96 % (95 % CI 93-99 %) were detected by blood and bone marrow cultures respectively. Blood culture sensitivity was 66 % (95 % CI 56-75 %) when compared with bone marrow culture results. The use of blood culture sensitivity as a proxy measure to estimate the proportion of typhoid fever cases detected by blood culture is likely to be an underestimate. As blood culture sensitivity is used as a correction factor in estimating typhoid disease burden, epidemiologists and policy makers should account for the underestimation. PMID:27188991

  20. Bio-oil upgrading strategies to improve PHA production from selected aerobic mixed cultures.

    PubMed

    Moita Fidalgo, Rita; Ortigueira, Joana; Freches, André; Pelica, João; Gonçalves, Magarida; Mendes, Benilde; Lemos, Paulo C

    2014-06-25

    Recent research on polyhydroxyalkanoates (PHAs) has focused on developing cost-effective production processes using low-value or industrial waste/surplus as substrate. One of such substrates is the liquid fraction resulting from pyrolysis processes, bio-oil. In this study, valorisation of bio-oil through PHA production was investigated. The impact of the complex bio-oil matrix on PHA production by an enriched mixed culture was examined. The performance of the direct utilization of pure bio-oil was compared with the utilization of three defined substrates contained in this bio-oil: acetate, glucose and xylose. When compared with acetate, bio-oil revealed lower capacity for polymer production as a result of a lower polymer yield on substrate and a lower PHA cell content. Two strategies for bio-oil upgrade were performed, anaerobic fermentation and vacuum distillation, and the resulting liquid streams were tested for polymer production. The first one was enriched in volatile fatty acids and the second one mainly on phenolic and long-chain fatty acids. PHA accumulation assays using the upgraded bio-oils attained polymer yields on substrate similar or higher than the one achieved with acetate, although with a lower PHA content. The capacity to use the enriched fractions for polymer production has yet to be optimized. The anaerobic digestion of bio-oil could also open-up the possibility to use the fermented bio-oil directly in the enrichment process of the mixed culture. This would increase the selective pressure toward an optimized PHA accumulating culture selection. PMID:24189432

  1. Preferential Use of Carbon Sources in Culturable Aerobic Mesophilic Bacteria of Coptotermes curvignathus's (Isoptera: Rhinotermitidae) Gut and Its Foraging Area.

    PubMed

    Wong, W Z; H'ng, P S; Chin, K L; Sajap, Ahmad Said; Tan, G H; Paridah, M T; Othman, Soni; Chai, E W; Go, W Z

    2015-10-01

    The lower termite, Coptotermes curvignathus, is one of the most prominent plantation pests that feed upon, digest, and receive nourishment from exclusive lignocellulose diets. The objective of this study was to examine the utilization of sole carbon sources by isolated culturable aerobic bacteria among communities from the gut and foraging pathway of C. curvignathus. We study the bacteria occurrence from the gut of C. curvignathus and its surrounding feeding area by comparing the obtained phenotypic fingerprint with Biolog's extensive species library. A total of 24 bacteria have been identified mainly from the family Enterobacteriaceae from the identification of Biolog Gen III. Overall, the bacteria species in the termite gut differ from those of foraging pathway within a location, except Acintobacter baumannii, which was the only bacteria species found in both habitats. Although termites from a different study area do not have the same species of bacteria in the gut, they do have a bacterial community with similar role in degrading certain carbon sources. Sugars were preferential in termite gut isolates, while nitrogen carbon sources were preferential in foraging pathway isolates. The preferential use of specific carbon sources by these two bacterial communities reflects the role of bacteria for regulation of carbon metabolism in the termite gut and foraging pathway. PMID:26314017

  2. Recalcitrance of 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (DDE) to cometabolic degradation by pure cultures of aerobic and anaerobic bacteria.

    PubMed

    Megharaj, M; Jovcic, A; Boul, H L; Thiele, J H

    1997-08-01

    Pure cultures of aerobic and anaerobic bacteria capable of oxidation and reductive dehalogenation of chloroethylenes, and aerobic bacteria involved in biodegradation of polychlorinated biphenyls (PCBs) were screened for their ability to cometabolize the persistent pollutant 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (DDE). Bacterial cultures expressing methane monooxygenase (Methylosinus trichosporium), propane monooxygenase (Mycobacterium vaccae) and biphenyl 2,3-dioxygenase enzymes (Pseudomonas fluorescens and Rhodococcus globerulus), as well as bacteria reductively dechlorinating chloroethylenes (Acetobacterium woodii and Clostridium butyricum) could not degrade DDE. Cell-free extracts of M. trichosporium, M. vaccae, P. fluorescens and R. globerulus were also unable to transform DDE, indicating that cell wall and membrane diffusion barriers were not biodegradation limiting. These studies suggest that these bacteria can not degrade DDE, even when provided with cosubstrates that induce chlorophenyl- and dichloroethylene-group transforming enzymes. PMID:9294241

  3. Culturable diversity of aerobic halophilic archaea (Fam. Halobacteriaceae) from hypersaline, meromictic Transylvanian lakes.

    PubMed

    Baricz, Andreea; Cristea, Adorján; Muntean, Vasile; Teodosiu, Gabriela; Andrei, Adrian-Ştefan; Molnár, Imola; Alexe, Mircea; Rakosy-Tican, Elena; Banciu, Horia Leonard

    2015-03-01

    Perennially stratified salt lakes situated in the Transylvanian Basin (Central Romania) were surveyed for the diversity of culturable halophilic archaea (Fam. Halobacteriaceae). The physical and chemical characteristics of the waters indicated that all the investigated lakes were meromictic and neutral hypersaline. Samples collected from upper, intermediate, and deeper water layers and sediments were used for the isolation of halophilic strains followed by 16S rRNA gene-based identification and phenotypic characterization. The phylogenetic analysis of the 16S rRNA gene sequences revealed that all 191 isolates reported in this study and 43 strains previously isolated were affiliated with the family Halobacteriaceae and classified to 18 genera. Haloferax was the most frequently isolated genus (~47 %), followed by Halobacterium spp. (~12 %), and Halorubrum spp. (~11 %). Highest culturable diversity was detected in Brâncoveanu Lake, the oldest and saltiest of all studied lakes, while the opposite was observed in the most stable and least human-impacted Fără Fund Lake. One strain from Ursu Lake might possibly constitute a novel Halorubrum species as shown by phylogenetic analysis. Several haloarchaeal taxa recently described in Asian (i.e., Iran, China) saline systems were also identified as inhabiting the Transylvanian salt lakes thus expanding our knowledege on the geographic distribution of Halobacteriaceae. PMID:25680859

  4. Blood Culture Bottle and Standard Culture Bottle Methods for Detection of Bacterial Pathogens in Parapneumonic Pleural Effusion

    PubMed Central

    Charoentunyarak, Surapan; Kananuraks, Sarassawan; Chindaprasirt, Jarin; Limpawattana, Panita; Sawanyawisuth, Kittisak

    2015-01-01

    Background: Bacterial parapneumonic pleural effusions (PPEs) have high morbidity. The accurate identification of pathogens is vital for initiating the appropriate treatment. A previous study suggested that the use of blood culture bottles might improve the bacterial yield in PPEs. Objectives: The aim of this study was to compare the culture positivity rate by the blood culture bottles and the standard culture bottles in bacterial PPEs. Patients and Methods: Patients diagnosed with PPEs at the Khon Kaen Hospital, Khon Kaen, Thailand, which is an endemic area of melioidosis, were enrolled consecutively and prospectively. The study period was from June first, 2012 to December 31st, 2013. The inclusion criteria were adult patients aged > 18 years, with exudative, neutrophilic parapneumonic effusion. Of the pleural fluid samples, 5 mL from all the eligible patients were collected in both blood culture bottles and the standard culture bottles. Patient baseline characteristics, laboratory results, and culture results were collected and analyzed. Results: During the study period, 129 patients met the study criteria. The bacteria-positive rate of pleural fluid culture using the standard culture bottle was 14.0%, whereas the positive rate using blood culture bottles was 24.0% (P < 0.001). Conclusions: The blood culture bottle method is more effective than the standard culture bottle method for the detection of bacterial pathogens in PPE. PMID:26587217

  5. Effect of Nordic Walking and Water Aerobics Training on Body Composition and the Blood Flow in Lower Extremities in Elderly Women

    PubMed Central

    Jasiński, Ryszard; Socha, Małgorzata; Sitko, Ludmiła; Kubicka, Katarzyna; Woźniewski, Marek; Sobiech, Krzysztof A.

    2015-01-01

    Nordic walking and water aerobics are very popular forms of physical activity in the elderly population. The aim of the study was to evaluate the influence of regular health training on the venous blood flow in lower extremities and body composition in women over 50 years old. Twenty-four women of mean age 57.9 (± 3.43) years, randomly divided into three groups (Nordic walking, water aerobics, and non-training), participated in the study. The training lasted 8 weeks, with one-hour sessions twice a week. Dietary habits were not changed. Before and after training vein refilling time and the function of the venous pump of the lower extremities were measured by photoplethysmography. Body composition was determined by bioelectrical impedance. Eight weeks of Nordic walking training improved the venous blood flow in lower extremities and normalized body composition in the direction of reducing chronic venous disorder risk factors. The average values of the refilling time variable (p = 0.04, p = 0.02, respectively) decreased in both the right and the left leg. After training a statistically significant increase in the venous pump function index was found only in the right leg (p = 0.04). A significant increase in fat-free mass, body cell mass and total body water was observed (p = 0.01), whereas body mass, the body mass index, and body fat decreased (p < 0.03). With regard to water aerobic training, no similar changes in the functions of the venous system or body composition were observed. PMID:25964815

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

    PubMed

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

    2016-06-01

    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 < 0.01), abdominal surgery (p = 0.01), and hemodialysis (p = 0.02). In all bottles, 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 < 0.01). In the presence of 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

  7. Identification of Gram-Negative Bacteria and Genetic Resistance Determinants from Positive Blood Culture Broths by Use of the Verigene Gram-Negative Blood Culture Multiplex Microarray-Based Molecular Assay

    PubMed Central

    Ledeboer, Nathan A.; Lopansri, Bert K.; Dhiman, Neelam; Cavagnolo, Robert; Carroll, Karen C.; Granato, Paul; Thomson, Richard; Butler-Wu, Susan M.; Berger, Heather; Samuel, Linoj; Pancholi, Preeti; Swyers, Lettie; Hansen, Glen T.; Tran, Nam K.; Polage, Christopher R.; Thomson, Kenneth S.; Hanson, Nancy D.; Winegar, Richard

    2015-01-01

    Bloodstream infection is a serious condition associated with significant morbidity and mortality. The outcome of these infections can be positively affected by the early implementation of effective antibiotic therapy based on the identification of the infecting organism and genetic markers associated with antibiotic resistance. In this study, we evaluated the microarray-based Verigene Gram-negative blood culture (BC-GN) assay in the identification of 8 genus or species targets and 6 genetic resistance determinants in positive blood culture broths. A total of 1,847 blood cultures containing Gram-negative organisms were tested using the BC-GN assay. This comprised 729 prospective fresh, 781 prospective or retrospective frozen, and 337 simulated cultures representing 7 types of aerobic culture media. The results were compared to those with standard bacterial culture and biochemical identification with nucleic acid sequence confirmation of the resistance determinants. Among monomicrobial cultures, the positive percent agreement (PPA) of the BC-GN assay with the reference method was as follows; Escherichia coli, 100%; Klebsiella pneumoniae, 92.9%; Klebsiella oxytoca, 95.5%; Enterobacter spp., 99.3%; Pseudomonas aeruginosa, 98.9%; Proteus spp., 100%; Acinetobacter spp., 98.4%; and Citrobacter spp., 100%. All organism identification targets demonstrated >99.5% negative percent agreement (NPA) with the reference method. Of note, 25/26 cultures containing K. pneumoniae that were reported as not detected by the BC-GN assay were subsequently identified as Klebsiella variicola. The PPA for identification of resistance determinants was as follows; blaCTX-M, 98.9%; blaKPC, 100%; blaNDM, 96.2%; blaOXA, 94.3%; blaVIM, 100%; and blaIMP, 100%. All resistance determinant targets demonstrated >99.9% NPA. Among polymicrobial specimens, the BC-GN assay correctly identified at least one organism in 95.4% of the broths and correctly identified all organisms present in 54.5% of the broths

  8. Identification of Gram-Negative Bacteria and Genetic Resistance Determinants from Positive Blood Culture Broths by Use of the Verigene Gram-Negative Blood Culture Multiplex Microarray-Based Molecular Assay.

    PubMed

    Ledeboer, Nathan A; Lopansri, Bert K; Dhiman, Neelam; Cavagnolo, Robert; Carroll, Karen C; Granato, Paul; Thomson, Richard; Butler-Wu, Susan M; Berger, Heather; Samuel, Linoj; Pancholi, Preeti; Swyers, Lettie; Hansen, Glen T; Tran, Nam K; Polage, Christopher R; Thomson, Kenneth S; Hanson, Nancy D; Winegar, Richard; Buchan, Blake W

    2015-08-01

    Bloodstream infection is a serious condition associated with significant morbidity and mortality. The outcome of these infections can be positively affected by the early implementation of effective antibiotic therapy based on the identification of the infecting organism and genetic markers associated with antibiotic resistance. In this study, we evaluated the microarray-based Verigene Gram-negative blood culture (BC-GN) assay in the identification of 8 genus or species targets and 6 genetic resistance determinants in positive blood culture broths. A total of 1,847 blood cultures containing Gram-negative organisms were tested using the BC-GN assay. This comprised 729 prospective fresh, 781 prospective or retrospective frozen, and 337 simulated cultures representing 7 types of aerobic culture media. The results were compared to those with standard bacterial culture and biochemical identification with nucleic acid sequence confirmation of the resistance determinants. Among monomicrobial cultures, the positive percent agreement (PPA) of the BC-GN assay with the reference method was as follows; Escherichia coli, 100%; Klebsiella pneumoniae, 92.9%; Klebsiella oxytoca, 95.5%; Enterobacter spp., 99.3%; Pseudomonas aeruginosa, 98.9%; Proteus spp., 100%; Acinetobacter spp., 98.4%; and Citrobacter spp., 100%. All organism identification targets demonstrated >99.5% negative percent agreement (NPA) with the reference method. Of note, 25/26 cultures containing K. pneumoniae that were reported as not detected by the BC-GN assay were subsequently identified as Klebsiella variicola. The PPA for identification of resistance determinants was as follows; blaCTX-M, 98.9%; blaKPC, 100%; blaNDM, 96.2%; blaOXA, 94.3%; blaVIM, 100%; and blaIMP, 100%. All resistance determinant targets demonstrated >99.9% NPA. Among polymicrobial specimens, the BC-GN assay correctly identified at least one organism in 95.4% of the broths and correctly identified all organisms present in 54.5% of the broths

  9. Molecular identification of Candida orthopsilosis isolated from blood culture.

    PubMed

    Yong, P V C; Chong, P P; Lau, L Y; Yeoh, R S C; Jamal, F

    2008-02-01

    The incidence of candidemia and invasive candidiasis have increased markedly due to the increasing number of immunocompromised patients. There are five major medically important species of Candida with their frequency of isolation in the diminishing order namely Candida albicans, Candida parapsilosis, Candida tropicalis, Candida glabrata and Candida krusei. In addition, there are numerous other species of Candida which differ in their genetic makeup, virulence properties, drug susceptibilities and sugar assimilation capabilities. In this report, an unusual Candida species was isolated from the blood of two leukaemic patients. Conventional culture and biochemical tests identified the Candida species as C. parapsilosis. Using fungal-specific oligonucleotide primers ITS1 and ITS4, we managed to amplify the ribosomal RNA gene and its internal transcribed spacer region from the genomic DNA of these isolates. The PCR products were then purified and subjected to automated DNA sequencing using BLAST and CLUSTAL sequence analysis identified these isolates to be Candida orthopsilosis. Candida orthopsilosis is a new species recently identified in 2005, being morphologically indistinguishable from C. parapsilosis and was previously classified as a subspecies of C. parapsilosis. This report highlights the importance of complementing traditional culture and biochemical-based identification methods with DNA-based molecular assays such as PCR as the latter is more superior in terms of its discriminatory power and speed. PMID:18266075

  10. Enhancement of recovery of Neisseria meningitidis by gelatin in blood culture media.

    PubMed Central

    Pai, C H; Sorger, S

    1981-01-01

    The efficacy of gelatin for the recovery of Neisseria meningitidis from blood cultures was evaluated in a clinical setting. The organism was isolated from seven patients with meningococcal infections in blood culture media containing 1% gelatin. In contrast, only two blood cultures from these patients were positive in media without gelatin (P less than 0.05). Gelatin did not influence the recovery of other organisms isolated during this study. Conventional blood culture media may be supplemented with gelatin when meningococcemia is suspected. PMID:6790567

  11. Naturally Occurring Culturable Aerobic Gut Flora of Adult Phlebotomus papatasi, Vector of Leishmania major in the Old World

    PubMed Central

    Mukhopadhyay, Jaba; Braig, Henk R.; Rowton, Edgar D.; Ghosh, Kashinath

    2012-01-01

    Background Cutaneous leishmaniasis is a neglected, vector-borne parasitic disease and is responsible for persistent, often disfiguring lesions and other associated complications. Leishmania, causing zoonotic cutaneous leishmaniasis (ZCL) in the Old World are mainly transmitted by the predominant sand fly vector, Phlebotomus papatasi. To date, there is no efficient control measure or vaccine available for this widespread insect-borne infectious disease. Methodology/Principal Findings A survey was carried out to study the abundance of different natural gut flora in P. papatasi, with the long-term goal of generating a paratransgenic sand fly that can potentially block the development of Leishmania in the sand fly gut, thereby preventing transmission of leishmania in endemic disease foci. Sand flies, in particular, P. papatasi were captured from different habitats of various parts of the world. Gut microbes were cultured and identified using 16S ribosomal DNA analysis and a phylogenetic tree was constructed. We found variation in the species and abundance of gut flora in flies collected from different habitats. However, a few Gram-positive, nonpathogenic bacteria including Bacillus flexus and B. pumilus were common in most of the sites examined. Conclusion/Significance Our results indicate that there is a wide range of variation of aerobic gut flora inhabiting sand fly guts, which possibly reflect the ecological condition of the habitat where the fly breeds. Also, some species of bacteria (B. pumilus, and B. flexus) were found from most of the habitats. Important from an applied perspective of dissemination, our results support a link between oviposition induction and adult gut flora. PMID:22629302

  12. Degradation of Polycyclic Aromatic Hydrocarbons at Low Temperature under Aerobic and Nitrate-Reducing Conditions in Enrichment Cultures from Northern Soils

    PubMed Central

    Eriksson, Mikael; Sodersten, Erik; Yu, Zhongtang; Dalhammar, Gunnel; Mohn, William W.

    2003-01-01

    The potential for biodegradation of polycyclic aromatic hydrocarbons (PAHs) at low temperature and under anaerobic conditions is not well understood, but such biodegradation would be very useful for remediation of polluted sites. Biodegradation of a mixture of 11 different PAHs with two to five aromatic rings, each at a concentration of 10 μg/ml, was studied in enrichment cultures inoculated with samples of four northern soils. Under aerobic conditions, low temperature severely limited PAH biodegradation. After 90 days, aerobic cultures at 20°C removed 52 to 88% of the PAHs. The most extensive PAH degradation under aerobic conditions at 7°C, 53% removal, occurred in a culture from creosote-contaminated soil. Low temperature did not substantially limit PAH biodegradation under nitrate-reducing conditions. Under nitrate-reducing conditions, naphthalene, 2-methylnaphthalene, fluorene, and phenanthrene were degraded. The most extensive PAH degradation under nitrate-reducing conditions at 7°C, 39% removal, occurred in a culture from fuel-contaminated Arctic soil. In separate transfer cultures from the above Arctic soil, incubated anaerobically at 7°C, removal of 2-methylnaphthalene and fluorene was stoichiometrically coupled to nitrate removal. Ribosomal intergenic spacer analysis suggested that enrichment resulted in a few predominant bacterial populations, including members of the genera Acidovorax, Bordetella, Pseudomonas, Sphingomonas, and Variovorax. Predominant populations from different soils often included phylotypes with nearly identical partial 16S rRNA gene sequences (i.e., same genus) but never included phylotypes with identical ribosomal intergenic spacers (i.e., different species or subspecies). The composition of the enriched communities appeared to be more affected by presence of oxygen, than by temperature or source of the inoculum. PMID:12514005

  13. Differential Label-free Quantitative Proteomic Analysis of Shewanella oneidensis Cultured under Aerobic and Suboxic Conditions by Accurate Mass and Time Tag Approach

    SciTech Connect

    Fang, Ruihua; Elias, Dwayne A.; Monroe, Matthew E.; Shen, Yufeng; McIntosh, Martin; Wang, Pei; Goddard, Carrie D.; Callister, Stephen J.; Moore, Ronald J.; Gorby, Yuri A.; Adkins, Joshua N.; Fredrickson, Jim K.; Lipton, Mary S.; Smith, Richard D.

    2006-04-01

    We describe the application of liquid chromatography coupled to mass spectrometry (LC/MS) without the use of stable isotope labeling for differential quantitative proteomics analysis of whole cell lysates of Shewanella oneidensis MR-1 cultured under aerobic and sub-oxic conditions. Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) was used to initially identify peptide sequences, and LC coupled to Fourier transform ion cyclotron resonance mass spectrometry (LC-FTICR) was used to confirm these identifications, as well as measure relative peptide abundances. 2343 peptides, covering 668 proteins were identified with high confidence and quantified. Among these proteins, a subset of 56 changed significantly using statistical approaches such as SAM, while another subset of 56 that were annotated as performing housekeeping functions remained essentially unchanged in relative abundance. Numerous proteins involved in anaerobic energy metabolism exhibited up to a 10-fold increase in relative abundance when S. oneidensis is transitioned from aerobic to sub-oxic conditions.

  14. Vampires, Pasteur and reactive oxygen species. Is the switch from aerobic to anaerobic metabolism a preventive antioxidant defence in blood-feeding parasites?

    PubMed

    Oliveira, Pedro L; Oliveira, Marcus F

    2002-08-14

    Several species of parasites show a reduction of their respiratory activity along their developmental cycles after they start to feed on vertebrate blood, relying on anaerobic degradation of carbohydrates to achieve their energy requirements. Usually, these parasites choose not to breathe despite of living in an environment of high oxygen availability such as vertebrate blood. Absence of the 'Pasteur effect' in most of these parasites has been well documented. Interestingly, together with the switch from aerobic to anaerobic metabolism in these parasites, there is clear evidence pointing to an increase in their antioxidant defences. As the respiratory chain in mitochondria is a major site of production of reactive oxygen species (ROS), we propose here that the arrest of respiration constitutes an adaptation to avoid the toxic effects of ROS. This situation would be especially critical for blood-feeding parasites because ROS produced in mitochondria would interact with pro-oxidant products of blood digestion, such as haem and/or iron, and increase the oxidative damage to the parasite's cells. PMID:12163151

  15. All in the Blood: A Review of Aboriginal Australians' Cultural Beliefs About Blood and Implications for Biospecimen Research.

    PubMed

    Kowal, Emma; Greenwood, Ashley; McWhirter, Rebekah E

    2015-10-01

    Public participation in medical research and biobanking is considered key to advances in scientific discovery and translation to improved health care. Cultural concerns relating to blood have been found to affect the participation of indigenous peoples and minorities in research, but such concerns are rarely specified in the literature. This article presents a review of the role of blood in Australian Aboriginal cultures. We discuss the range of meanings and uses of blood in traditional culture, including their use in ceremonies, healing, and sorcery. We draw on more recent literature on Aboriginal Australians and biomedicine to consider how traditional beliefs may be changing over time. These findings provide an empirical basis for researchers and bioethicists to develop culturally grounded strategies to boost the participation of Aboriginal Australians in biomedical research. They also serve as a model for integrating anthropological literature with bioethical concerns that could be applied to other indigenous and minority groups. PMID:26376752

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

    PubMed

    Rahiman, Farzana; Pool, Edmund John

    2010-01-01

    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 < 0.001) and also the biomarker of humoral immunity, Interleukin-10 (P < 0.001). 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

  17. Growth and energy metabolism in aerobic fed-batch cultures of Saccharomyces cerevisiae: Simulation and model verification

    SciTech Connect

    Pham, H.T.B.; Larsson, G.; Enfors, S.O.

    1998-11-20

    Some yeast species are classified as being glucose sensitive, which means that they may produce ethanol also under aerobic conditions when the sugar concentration is high. A kinetic model of overflow metabolism in Saccharomyces cerevisiae was used for simulation of aerobic fed-batch cultivations. An inhibitory effect of ethanol on the maximum respiration of the yeast was observed in the experiments and included in the model. The model predicts respiration, biomass, and ethanol formation and the subsequent ethanol consumption, and was experimentally validated in fed-batch cultivations. Oscillating sugar feed with resulting oscillating carbon dioxide production did not influence the maximum respiration rate, which indicates that the pyruvate dehydrogenase complex is not involved as a bottleneck causing aerobic ethanol formation.

  18. Controlled Clinical Comparison of the BacT/ALERT FN and the Standard Anaerobic SN Blood Culture Medium

    PubMed Central

    Mirrett, S.; Petti, C. A.; Woods, C. W.; Magadia, R.; Weinstein, M. P.; Reller, L. B.

    2004-01-01

    To determine the optimal anaerobic companion bottle to pair with the BacT/ALERT (bioMérieux, Durham, N.C.) nonvented aerobic FA (FA) medium for recovery of pathogenic microorganisms from adult patients with bacteremia and fungemia, we compared the BacT/ALERT FN (FN) anaerobic bottle with the standard BacT/ALERT SN (SN) anaerobic bottle. Each bottle, FA, FN, and SN, was filled with 8 to 12 ml of blood. Of 11,498 blood culture sets received in the clinical microbiology laboratories at two university medical centers, 7,945 sets had all three bottles filled adequately and 8,569 had both anaerobic bottles filled adequately. Of 686 clinically important (based on previously published criteria) isolates detected in one or both adequately filled anaerobic bottles, more staphylococci (P < 0.001), including Staphylococcus aureus (P < 0.001); members of the family Enterobacteriaceae (P < 0.001); and all microorganisms combined (P < 0.001) were detected in FN bottles. In contrast, more Pseudomonas aeruginosa isolates (P < 0.01) and yeasts (P < 0.001) were detected in SN bottles. More Bacteroides fragilis group bacteremias were detected only in the FN (six) than in the SN (one) anaerobic bottle (P = not significant). Overall, the mean time to detection was shorter with FN (16.8 h) than with SN (18.2 h). This difference in time to detection was greatest for the B. fragilis group: FN, 28 h, versus SN, 60.0 h. Many of the facultative microorganisms recovered in either FN or SN were also found in the companion FA. When microorganisms found in the companion FA bottle were omitted from the analysis, significantly more staphylococci (P < 0.001), including S. aureus (P < 0.001), and Enterobacteriaceae (P < 0.005) still were detected in FN bottles, whereas there were no significant differences for P. aeruginosa and yeasts, which were found as expected in FA bottles. We conclude that the companion anaerobic FN bottle detects more microorganisms than does the anaerobic SN bottle when used

  19. PCR amplification of Bartonella koehlerae from human blood and enrichment blood cultures

    PubMed Central

    2010-01-01

    Background Cats appear to be the primary reservoir host for Bartonella koehlerae, an alpha Proteobacteria that is most likely transmitted among cat populations by fleas (Ctenocephalides felis). Bartonella koehlerae has caused endocarditis in a dog and in one human patient from Israel, but other clinically relevant reports involving this bacterium are lacking. Despite publication of numerous, worldwide epidemiological studies designed to determine the prevalence of Bartonella spp. bacteremia in cats, B. koehlerae has never been isolated using conventional blood agar plates. To date, successful isolation of B. koehlerae from cats and from the one human endocarditis patient has consistently required the use of chocolate agar plates. Results In this study, Bartonella koehlerae bacteremia was documented in eight immunocompetent patients by PCR amplification and DNA sequencing, either prior to or after enrichment blood culture using Bartonella alpha Proteobacteria growth medium. Presenting symptoms most often included fatigue, insomnia, joint pain, headache, memory loss, and muscle pain. Four patients were also infected with Bartonella vinsonii subsp. berkhoffii genotype II. After molecular documentation of B. koehlerae infection in these patients, a serological test was developed and serum samples were tested retrospectively. Bartonella koehlerae antibodies were not detected (titers < 1:16) in 30 healthy human control sera, whereas five of eight patient samples had B. koehlerae antibody titers of 1:64 or greater. Conclusions Although biased by a study population consisting of individuals with extensive arthropod and animal exposure, the results of this study suggest that B. koehlerae bacteremia is more common in immunocompetent people than has been previously suspected. Future studies should more thoroughly define modes of transmission and risk factors for acquiring infection with B. koehlerae. In addition, studies are needed to determine if B. koehlerae is a cause or

  20. Inhibition of nitric oxide and prostaglandins, but not endothelial-derived hyperpolarizing factors, reduces blood flow and aerobic energy turnover in the exercising human leg.

    PubMed

    Mortensen, Stefan P; González-Alonso, José; Damsgaard, Rasmus; Saltin, Bengt; Hellsten, Ylva

    2007-06-01

    Prostaglandins, nitric oxide (NO) and endothelial-derived hyperpolarizing factors (EDHFs) are substances that have been proposed to be involved in the regulation of skeletal muscle blood flow during physical activity. We measured haemodynamics, plasma ATP at rest and during one-legged knee-extensor exercise (19 +/- 1 W) in nine healthy subjects with and without intra-arterial infusion of indomethacin (Indo; 621 +/- 17 microg min(-1)), Indo + N(G)-monomethyl-L-arginine (L-NMMA; 12.4 +/- 0.3 mg min(-1)) (double blockade) and Indo + L-NMMA + tetraethylammonium chloride (TEA; 12.4 +/- 0.3 mg min(-1)) (triple blockade). Double and triple blockade lowered leg blood flow (LBF) at rest (P<0.05), while it remained unchanged with Indo. During exercise, LBF and vascular conductance were 2.54 +/- 0.10 l min(-1) and 25 +/- 1 mmHg, respectively, in control and they were lower with double (33 +/- 3 and 36 +/- 4%, respectively) and triple (26 +/- 4 and 28 +/- 3%, respectively) blockade (P<0.05), while there was no difference with Indo. The lower LBF and vascular conductance with double and triple blockade occurred in parallel with a lower O(2) delivery, cardiac output, heart rate and plasma [noradrenaline] (P<0.05), while blood pressure remained unchanged and O(2) extraction and femoral venous plasma [ATP] increased. Despite the increased O(2) extraction, leg was 13 and 17% (triple and double blockade, respectively) lower than control in parallel to a lower femoral venous temperature and lactate release (P<0.05). These results suggest that NO and prostaglandins play important roles in skeletal muscle blood flow regulation during moderate intensity exercise and that EDHFs do not compensate for the impaired formation of NO and prostaglandins. Moreover, inhibition of NO and prostaglandin formation is associated with a lower aerobic energy turnover and increased concentration of vasoactive ATP in plasma. PMID:17347273

  1. Culture-Independent Analysis of Bacterial Fuel Contamination Provides Insight into the Level of Concordance with the Standard Industry Practice of Aerobic Cultivation ▿ †

    PubMed Central

    White, Judith; Gilbert, Jack; Hill, Graham; Hill, Edward; Huse, Susan M.; Weightman, Andrew J.; Mahenthiralingam, Eshwar

    2011-01-01

    Bacterial diversity in contaminated fuels has not been systematically investigated using cultivation-independent methods. The fuel industry relies on phenotypic cultivation-based contaminant identification, which may lack accuracy and neglect difficult-to-culture taxa. By the use of industry practice aerobic cultivation, 16S rRNA gene sequencing, and strain genotyping, a collection of 152 unique contaminant isolates from 54 fuel samples was assembled, and a dominance of Pseudomonas (21%), Burkholderia (7%), and Bacillus (7%) was demonstrated. Denaturing gradient gel electrophoresis (DGGE) of 15 samples revealed Proteobacteria and Firmicutes to be the most abundant phyla. When 16S rRNA V6 gene pyrosequencing of four selected fuel samples (indicated by “JW”) was performed, Betaproteobacteria (42.8%) and Gammaproteobacteria (30.6%) formed the largest proportion of reads; the most abundant genera were Marinobacter (15.4%; JW57), Achromobacter (41.6%; JW63), Burkholderia (80.7%; JW76), and Halomonas (66.2%; JW78), all of which were also observed by DGGE. However, the Clostridia (38.5%) and Deltaproteobacteria (11.1%) identified by pyrosequencing in sample JW57 were not observed by DGGE or aerobic culture. Genotyping revealed three instances where identical strains were found: (i) a Pseudomonas sp. strain recovered from 2 different diesel fuel tanks at a single industrial site; (ii) a Mangroveibacter sp. strain isolated from 3 biodiesel tanks at a single refinery site; and (iii) a Burkholderia vietnamiensis strain present in two unrelated automotive diesel samples. Overall, aerobic cultivation of fuel contaminants recovered isolates broadly representative of the phyla and classes present but lacked accuracy by overrepresenting members of certain groups such as Pseudomonas. PMID:21602386

  2. Culture-independent analysis of bacterial fuel contamination provides insight into the level of concordance with the standard industry practice of aerobic cultivation.

    PubMed

    White, Judith; Gilbert, Jack; Hill, Graham; Hill, Edward; Huse, Susan M; Weightman, Andrew J; Mahenthiralingam, Eshwar

    2011-07-01

    Bacterial diversity in contaminated fuels has not been systematically investigated using cultivation-independent methods. The fuel industry relies on phenotypic cultivation-based contaminant identification, which may lack accuracy and neglect difficult-to-culture taxa. By the use of industry practice aerobic cultivation, 16S rRNA gene sequencing, and strain genotyping, a collection of 152 unique contaminant isolates from 54 fuel samples was assembled, and a dominance of Pseudomonas (21%), Burkholderia (7%), and Bacillus (7%) was demonstrated. Denaturing gradient gel electrophoresis (DGGE) of 15 samples revealed Proteobacteria and Firmicutes to be the most abundant phyla. When 16S rRNA V6 gene pyrosequencing of four selected fuel samples (indicated by "JW") was performed, Betaproteobacteria (42.8%) and Gammaproteobacteria (30.6%) formed the largest proportion of reads; the most abundant genera were Marinobacter (15.4%; JW57), Achromobacter (41.6%; JW63), Burkholderia (80.7%; JW76), and Halomonas (66.2%; JW78), all of which were also observed by DGGE. However, the Clostridia (38.5%) and Deltaproteobacteria (11.1%) identified by pyrosequencing in sample JW57 were not observed by DGGE or aerobic culture. Genotyping revealed three instances where identical strains were found: (i) a Pseudomonas sp. strain recovered from 2 different diesel fuel tanks at a single industrial site; (ii) a Mangroveibacter sp. strain isolated from 3 biodiesel tanks at a single refinery site; and (iii) a Burkholderia vietnamiensis strain present in two unrelated automotive diesel samples. Overall, aerobic cultivation of fuel contaminants recovered isolates broadly representative of the phyla and classes present but lacked accuracy by overrepresenting members of certain groups such as Pseudomonas. PMID:21602386

  3. Evaluation of PNA FISH® Yeast Traffic Light in identification of Candida species from blood and non-blood culture specimens.

    PubMed

    Radic, Marina; Goic-Barisic, Ivana; Novak, Anita; Rubic, Zana; Tonkic, Marija

    2016-08-01

    PNA FISH(®) (peptide nucleic acid fluorescent in situ hybridization) Yeast Traffic Light (PNA FISH(®) YTL) assay is a commercially avaliable method for rapid identification of Candida spp. directly from positive blood cultures. This report provides a one-year experience in identification of yeasts from 25 specimens (15 positive blood cultures and 10 other clinically significant specimens) using PNA FISH(®) YTL and comparing it to VITEK 2 System. Overall, assay identification compatibility with VITEK 2 System was found among 21/25 (84%) isolates tested. Only 3/25 (12%) of the isolates were not identified, and one isolate was misidentified by the PNA FISH(®) YTL assay. Our results show that the assay is a reliable method in identification of Candida spp. not only from blood cultures, but even from other clinically significant specimens (urine cultures, catheter tip cultures, peritoneal fluid cultures) when compared to automated method like VITEK 2 System. This novel application of the PNA FISH(®) YTL assay could therefore contribute to cost savings and significant benefit to patients, as rapid information about isolated yeast species is provided. PMID:27067303

  4. Effects of moderate-intensity aerobic cycling and swim exercise on post-exertional blood pressure in healthy young untrained and triathlon-trained men and women.

    PubMed

    Lakin, Robert; Notarius, Catherine; Thomas, Scott; Goodman, Jack

    2013-12-01

    Aerobic exercises such as running, walking and cycling are known to elicit a PEH (post-exercise hypotensive) response in both trained and UT (untrained) subjects. However, it is not known whether swim exercise produces a similar effect in normotensive individuals. The complex acute physiological responses to water immersion suggest swimming may affect BP (blood pressure) differently than other forms of aerobic exercises. We tested the hypothesis that an acute bout of swimming would fail to elicit a PEH BP response compared with an equivalent bout of stationary cycling, regardless of training state. We studied 11 UT and ten triathlon-trained young healthy normotensive [SBP/DBP (systolic BP/diastolic BP) <120/80 mmHg)] men and women (age 23±1 years) who underwent 30 min of intensity-matched cycling and swimming sessions to assess changes in BP during a 75-min seated recovery. CO (cardiac output), SV (stroke volume), TPR (total peripheral resistance), HR (heart rate), HRV (HR variability) and core and skin temperature were also assessed. In UT subjects, PEH was similar between cycling (-3.1±1 mmHg) and swimming (-5.8±1 mmHg), with the greater magnitude of PEH following swimming, reflecting a significant fall in SV between modalities (P<0.05). Trained individuals did not exhibit a PEH response following swimming (0.3±1 mmHg), yet had a significant fall in SBP at 50 min post-cycling exercise (-3.7±1 mmHg) (P<0.05). The absence of PEH after swimming in the trained group may reflect a higher cardiac sympathetic outflow [as indicated by the LF (low-frequency) spectral component of HRV) (25 and 50 min) (P<0.05)] and a slower return of vagal tone, consistent with a significant increase in HR between modalities at all time points (P<0.05). These results suggest that training may limit the potential for an effective post-exertional hypotensive response to aerobic swimming. PMID:23763298

  5. Independent influence of negative blood cultures and bloodstream infections on in-hospital mortality

    PubMed Central

    2014-01-01

    Background The independent influence of blood culture testing and bloodstream infection (BSI) on hospital mortality is unclear. Methods We included all adults treated in non-psychiatric services at our hospital between 2004 and 2011. We identified all blood cultures and their results to determine the independent association of blood culture testing and BSI on death in hospital using proportional hazards modeling that adjusted for important covariates. Results Of 297 070 hospitalizations, 48 423 had negative blood cultures and 5274 had BSI. 12 529 (4.2%) died in hospital. Compared to those without blood cultures, culture-negative patients and those with BSI were sicker. Culture-negative patients had a significantly increased risk of death in hospital (adjusted hazard ratio [HR] ranging between 3.1 and 4.4 depending on admission urgency, extent of comorbidities, and whether the blood culture was taken in the intensive care unit). Patients with BSI had a significantly increased risk of death (adj-HR ranging between 3.8 and 24.3] that was significantly higher when BSI was: diagnosed within the first hospital day; polymicrobial; in patients who were exposed to immunosuppressants or were neutropenic; or due to Clostridial and Candidal organisms. Death risk in culture negative and bloodstream infection patients decreased significantly with time. Conclusions Risk of death in hospital is independently increased both in patients with negative blood cultures and further in those with bloodstream infection. Death risk associated with bloodstream infections varied by the patient’s immune status and the causative microorganism. PMID:24444097

  6. Is a single positive blood culture for Enterococcus species representative of infection or contamination?

    PubMed

    Jindai, K; Strerath, M S; Hess, T; Safdar, N

    2014-11-01

    Data on the clinical outcomes of patients with a single compared with multiple positive blood cultures for Enterococcus species is limited. We undertook a retrospective cohort study in adults with at least one positive blood culture for Enterococcus species in a single institution. Clinical outcomes included death and elimination of infection. We included 471 positive blood cultures from 206 enterococcal positive blood culture episodes in 189 patients. Multiple positive blood cultures for Enterococcus species occurred in 110/206 (53.4 %) episodes; 31.6 % of patients had diabetes mellitus; 42.9 % of patients had solid or hematologic malignancy; 26.5 % of patients were solid organ transplant recipients; hospital-acquired and healthcare-associated acquisition represented 55.3 % and 33.0 % of episodes, respectively. Thirty-five patients died and 110 episodes of enterococcal bloodstream infection were successfully treated. In the multivariable analysis, multiple positive blood cultures were not statistically significantly associated with an increased likelihood of in-hospital death [odds ratio (OR) 1.00, 95 % confidence interval (CI) 0.42-2.40] or elimination (OR 1.41, 95 % CI 0.76-2.64) compared with single positive blood cultures. Hematologic malignancy and diabetes mellitus were independently associated with in-hospital death (OR 2.83, 95 % Cl 1.02-7.82; OR 2.79, 95 % Cl 1.16-6.70, respectively). Infectious disease consultation was associated with a greater likelihood of elimination (OR 2.50, 95 % Cl 1.32-4.72). The clinical outcomes of patients with single versus multiple positive blood cultures with Enterococcus species were similar in our institution. Further studies should examine efficient methods to detect contamination versus true infection. PMID:25027071

  7. Tumor necrosis factor-alpha (TNF-alpha) concentrations from whole blood cultures correlate with isolated peripheral blood mononuclear cell cultures

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Many cellular immune assays are impractical because they require labor-intensive isolation of cells from their natural environment. The objectives of this study were to determine the relationship between cell culture supernatant TNF-alpha from isolated peripheral blood mononuclear cells (PBMC) and w...

  8. MALDI-TOF mass spectrometry for early identification of bacteria grown in blood culture bottles.

    PubMed

    Zabbe, Jean-Benoît; Zanardo, Laura; Mégraud, Francis; Bessède, Emilie

    2015-08-01

    This note reports an interesting way to rapidly identify bacteria grown from blood culture bottles. Chocolate agar plates were inoculated with 1 drop of the positive blood bottle medium. After a 3-hour incubation, the growth veil was submitted to MALDI-TOF mass spectrometry: 77% of the bacteria present have been correctly identified. PMID:25940929

  9. Direct testing of blood culture for detection of the serotype 5 and 8 capsular polysaccharides of Staphylococcus aureus.

    PubMed

    Boutonnier, A; Nato, F; Bouvet, A; Lebrun, L; Audurier, A; Mazie, J C; Fournier, J M

    1989-05-01

    Monoclonal antibodies (MAbs) reactive with serotype 5 and 8 capsular polysaccharides of Staphylococcus aureus have been used to test, by enzyme-linked immunosorbent assay (ELISA), blood culture fluids for the presence of S. aureus. A total of 748 blood cultures from 665 patients yielding 706 bacterial isolates belonging to more than 26 bacterial species were studied. All blood cultures containing bacterial strains belonging to species other than S. aureus were negative in ELISA. All 23 blood cultures containing serotype 5 S. aureus were positive in ELISA with the corresponding MAb. Out of 20 blood cultures containing serotype 8 S. aureus, 19 were positive with the corresponding MAb. All 5 blood cultures containing nontypeable S. aureus were negative in ELISA with both MAbs. This method provides reliable identification of serotype 5 or serotype 8 S. aureus by direct testing of blood culture fluids with ELISA. PMID:2745705

  10. Direct testing of blood culture for detection of the serotype 5 and 8 capsular polysaccharides of Staphylococcus aureus.

    PubMed Central

    Boutonnier, A; Nato, F; Bouvet, A; Lebrun, L; Audurier, A; Mazie, J C; Fournier, J M

    1989-01-01

    Monoclonal antibodies (MAbs) reactive with serotype 5 and 8 capsular polysaccharides of Staphylococcus aureus have been used to test, by enzyme-linked immunosorbent assay (ELISA), blood culture fluids for the presence of S. aureus. A total of 748 blood cultures from 665 patients yielding 706 bacterial isolates belonging to more than 26 bacterial species were studied. All blood cultures containing bacterial strains belonging to species other than S. aureus were negative in ELISA. All 23 blood cultures containing serotype 5 S. aureus were positive in ELISA with the corresponding MAb. Out of 20 blood cultures containing serotype 8 S. aureus, 19 were positive with the corresponding MAb. All 5 blood cultures containing nontypeable S. aureus were negative in ELISA with both MAbs. This method provides reliable identification of serotype 5 or serotype 8 S. aureus by direct testing of blood culture fluids with ELISA. PMID:2745705

  11. Controlled comparison of the BacT/Alert and BACTEC 660/730 nonradiometric blood culture systems.

    PubMed Central

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

    1992-01-01

    In a collaborative study at three university hospitals, the recovery of microorganisms and the speed of detection of microbial growth by the BacT/Alert (Organon Teknika Corporation, Durham, N.C.) and BACTEC 660/730 (Becton-Dickinson Diagnostic Instrument Systems, Sparks, Md.) nonradiometric blood culture systems were compared. A total of 5,918 comparisons were made between BacT/Alert aerobic and BACTEC NR 6A bottles and 5,992 comparisons were made between BacT/Alert anaerobic and BACTEC NR 7A bottles. Each bottle was inoculated with 5 ml of blood. The overall recoveries of microorganisms from the two aerobic bottles were comparable; members of the family Enterobacteriaceae were recovered more often from BacT/Alert aerobic bottles alone (P less than 0.001). The overall recoveries of microorganisms from the anaerobic bottles were not significantly different. Growth of Staphylococcus aureus (P less than 0.001), coagulase-negative staphylococci (P less than 0.01), streptococci (P less than 0.001), Escherichia coli (P less than 0.01), other members of the family Enterobacteriaceae (P less than 0.02), and Pseudomonas aeruginosa (P less than 0.05) was detected earlier in BacT/Alert aerobic bottles. Growth of S. aureus (P less than 0.001), coagulase-negative staphylococci (P less than 0.05), enterococci (P less than 0.01), Streptococcus pneumoniae (P less than 0.02), viridans group streptococci (P less than 0.05), E. coli (P less than 0.001), Klebsiella pneumoniae (P less than 0.01), and other members of the family Enterobacteriaceae (P less than 0.001) was detected earlier in BacT/Alert anaerobic bottles. In a system-versus-system comparison, more gram-positive cocci were recovered from the BACTEC system alone (P < 0.05), and more members or the family Enterobacteriaceae were recovered from the BacT/Alert system alone (P < 0.001). As a system, the BacT/Alert system detected growth of S. aureus (P < 0.001), coagulase-negative staphylococci (P < 0.01), streptococci (P < 0

  12. Controlled clinical evaluation of BACTEC Plus Aerobic/F and BacT/Alert Aerobic FAN bottles for detection of bloodstream infections.

    PubMed Central

    Pohlman, J K; Kirkley, B A; Easley, K A; Basille, B A; Washington, J A

    1995-01-01

    A total of 7,190 blood culture sets were obtained from adult patients with a suspected bloodstream infection. A 20-ml sample of blood was distributed equally between the aerobic FAN bottle which was monitored in the BacT/Alert system and a Plus Aerobic/F bottle which was monitored in the BACTEC 9240 system. A total of 988 positive cultures were obtained from 483 patients; however, only 453 positive cultures from 173 patients met the criteria for volume ( > or = ml per bottle) and clinical significance on the basis of concurrent case review required for data analysis. There were 25 and 68 false positives from the FAN and Plus Aerobic/F bottles, respectively. There were no statistically significant differences between systems in the number of positive cultures or septic episodes by species; however, the total number of Enterobacteriaceae and Pseudomonas aeruginosa isolates combined was significantly greater in the FAN bottle (P = 0.04). Detection times did not differ significantly between systems for positive cultures; however, episodes of Staphylococcus aureus bacteremia were detected significantly more rapidly from the FAN bottle (P = 0.005). There was no significant difference between systems in the detection of bloodstream infections in patients receiving antibiotics at the time of blood culture. PMID:8576333

  13. Hostility, cultural orientation, and casual blood pressure readings in African Americans.

    PubMed

    Daniels, I N; Harrell, J P; Floyd, L J; Bell, S R

    2001-01-01

    Evidence suggests that hostility correlates with blood pressure levels in African-American samples. However, some studies have reported an inverse relationship, while others have found the relationship between blood pressure and hostility to be positive. Other literature suggests health outcomes in general, and blood pressure in particular, are related to cultural orientation in African-American samples. In the present study, six casual measures of blood pressure and heart rate in a sample of 90 African-American college students were aggregated and correlated with measures of hostility and cultural orientation. Correlational and regression analyses revealed a weak positive relationship between hostility and systolic blood pressure. The relationships between the cardiovascular measures and cultural orientation were more consistent. The tendency to embrace mainstream Euro-American values, such as materialism, individuality, and competitiveness, was associated with more rapid heart rate and higher diastolic blood pressure levels for both men and women. The relationship between systolic blood pressure and cultural orientation emerged for men only. The findings encourage further research into the relationship between personality variables and cardiovascular activity in African-American samples. PMID:11763302

  14. Comparison of length of stay and outcomes of patients with positive versus negative blood culture results

    PubMed Central

    Hozhabri, Neda S. T.; Armstrong, Kris; Puthottile, Jason; Benavides, Raul; Beal, Stacy

    2015-01-01

    In the United States, sepsis is the leading cause of death in critically ill patients. The fatality rate for severe sepsis is about 40%, and treatment costs over $16 billion annually. It is critical to identify and treat the source of sepsis. While there are varying guidelines determining when to draw blood for culture, at Baylor University Medical Center at Dallas, blood cultures are ordered for patients with new onset of fever, immunosuppression, or a suspicion of an underlying infectious etiology. We conducted a retrospective study of patients who had blood cultures after hospital admission or in the emergency department in December 2013. We compared length of stay and outcomes of patients with positive versus negative blood cultures. There was no significant difference for length of stay or outcomes among patients with positive and negative blood cultures. For patients admitted from the emergency department, there was a longer length of stay for patients with positive cultures; however, the overall prognosis was not worse. PMID:25552786

  15. Use of prototype automated blood culture system and gas-liquid chromatography for the analysis of continuous ambulatory peritoneal dialysis associated infection.

    PubMed Central

    Catchpole, C R; Macrae, F; Brown, J D; Palmer, M; Healing, D E; Richards, N T; Elliott, T S

    1997-01-01

    AIMS: (1) To compare the recovery of organisms from continuous ambulatory peritoneal dialysis (CAPD) effluent fluid obtained from patients with clinical evidence of peritonitis, with an automated system (AS) and the Septichek blood culture system; (2) to evaluate the times to detection of organisms with the two systems; (3) to identify anaerobes from CAPD samples by extended anaerobic culture and gas-liquid chromatography (GLC). METHODS: 168 CAPD effluent fluid samples were studied, representing 157 episodes of peritonitis in 97 patients. CAPD samples were inoculated into two AS bottles-one anaerobic, one aerobic-and a Septichek bottle; samples were also examined for cell count, Gram stain, and direct culture. Culture bottles were then subcultured onto various media, and any organisms isolated were identified. After routine culture, GLC was performed on culture fluid in the anaerobic AS and Septichek bottles. When volatile fatty acids were detected, the broths were cultured anaerobically on specialised medium for a further five days. RESULTS: 147 organisms were isolated from the 168 samples: 96 (57%) yielded growth of significant organisms by direct culture, as compared to 129 (76.8%) by both AS and Septichek. There was no significant difference in isolation rates between AS and Septichek, but time to detection was more rapid with the AS system (p < 0.002). GLC showed volatile fatty acid in 15 specimens; of these, 14 subsequently grew anaerobic organisms. CONCLUSIONS: AS was comparable to Septichek for numbers of isolations. Speed to detection was faster with the AS, which may be an advantage in management of patients with CAPD peritonitis. GLC showed anaerobes in several cases which would not have been detected without prolonged anaerobic culture; thus anaerobic cultures are recommended for patients who are unresponsive to antimicrobials or who have evidence of bowel perforation. PMID:9155676

  16. Reconstitution activity of hypoxic cultured human cord blood CD34-positive cells in NOG mice

    SciTech Connect

    Shima, Haruko; Takubo, Keiyo; Iwasaki, Hiroko; Yoshihara, Hiroki; Gomei, Yumiko; Hosokawa, Kentaro; Arai, Fumio; Takahashi, Takao; Suda, Toshio

    2009-01-16

    Hematopoietic stem cells (HSCs) reside in hypoxic areas of the bone marrow. However, the role of hypoxia in the maintenance of HSCs has not been fully characterized. We performed xenotransplantation of human cord blood cells cultured in hypoxic or normoxic conditions into adult NOD/SCID/IL-2R{gamma}{sup null} (NOG) mice. Hypoxic culture (1% O{sub 2}) for 6 days efficiently supported the maintenance of HSCs, although cell proliferation was suppressed compared to the normoxic culture. In contrast, hypoxia did not affect in vitro colony-forming ability. Upregulation of a cell cycle inhibitor, p21, was observed in hypoxic culture. Immunohistochemical analysis of recipient bone marrow revealed that engrafted CD34{sup +}CD38{sup -} cord blood HSCs were hypoxic. Taken together, these results demonstrate the significance of hypoxia in the maintenance of quiescent human cord blood HSCs.

  17. Evaluation of Three Rapid Diagnostic Methods for Direct Identification of Microorganisms in Positive Blood Cultures

    PubMed Central

    Martinez, Raquel M.; Bauerle, Elizabeth R.; Fang, Ferric C.

    2014-01-01

    The identification of organisms from positive blood cultures generally takes several days. However, recently developed rapid diagnostic methods offer the potential for organism identification within only a few hours of blood culture positivity. In this study, we evaluated the performance of three commercial methods to rapidly identify organisms directly from positive blood cultures: QuickFISH (AdvanDx, Wolburn, MA), Verigene Gram-Positive Blood Culture (BC-GP; Nanosphere, Northbrook, IL), and matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) with Sepsityper processing (Bruker Daltonics, Billerica, MA). A total of 159 blood cultures (VersaTREK Trek Diagnostic Systems, Cleveland, OH) positive for Gram-positive and Gram-negative bacteria as well as yeast were analyzed with QuickFISH and MALDI-TOF MS. In all, 102 blood cultures were analyzed using the BC-GP assay. For monomicrobial cultures, we observed 98.0% concordance with routine methods for both QuickFISH (143/146) and the BC-GP assay (93/95). MALDI-TOF MS demonstrated 80.1% (117/146) and 87.7% (128/146) concordance with routine methods to the genus and species levels, respectively. None of the methods tested were capable of consistently identifying polymicrobial cultures in their entirety or reliably differentiating Streptococcus pneumoniae from viridans streptococci. Nevertheless, the methods evaluated in this study are convenient and accurate for the most commonly encountered pathogens and have the potential to dramatically reduce turnaround time for the provision of results to the treating physician. PMID:24808235

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

    PubMed

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

    1993-01-01

    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

  19. Lysophosphatidic acid does not cause blood/lymphatic vessel plasticity in the rat mesentery culture model.

    PubMed

    Sweat, Richard S; Azimi, Mohammad S; Suarez-Martinez, Ariana D; Katakam, Prasad; Murfee, Walter L

    2016-07-01

    Understanding the mechanisms behind endothelial cell identity is crucial for the goal of manipulating microvascular networks. Lysophosphatidic acid (LPA) and serum stimulation have been suggested to induce a lymphatic identity in blood endothelial cells in vitro. The objective of this study was to determine if LPA or serum induces blood-to-lymphatic vessel phenotypic transition in microvascular networks. The rat mesentery culture model was used to observe the effect of stimulation on blood and lymphatic microvascular networks ex vivo. Vascularized mesenteric tissues were harvested from adult Wistar rats and cultured with LPA or 10% serum for up to 5 days. Tissues were then immunolabeled with PECAM to identify blood vessels and LYVE-1 or Prox1 to identify lymphatic vessels. We show that while LPA caused capillary sprouting and increased vascular length density in adult microvascular networks, LPA did not cause a blood-to-lymphatic phenotypic transition. The results suggest that LPA is not sufficient to cause blood endothelial cells to adopt a lymphatic identity in adult microvascular networks. Similarly, serum stimulation caused robust angiogenesis and increased lymphatic/blood vessel connections, yet did not induce a blood-to-lymphatic phenotypic transition. Our study highlights an understudied area of lymphatic research and warrants future investigation into the mechanisms responsible for the maintenance of blood and lymphatic vessel identity. PMID:27401461

  20. Fluorescent In Situ Hybridization Allows Rapid Identification of Microorganisms in Blood Cultures

    PubMed Central

    Kempf, Volkhard A. J.; Trebesius, Karlheinz; Autenrieth, Ingo B.

    2000-01-01

    Using fluorescent in situ hybridization (FISH) with rRNA-targeted fluorescently labelled oligonucleotide probes, pathogens were rapidly detected and identified in positive blood culture bottles without cultivation and biotyping. In this study, 115 blood cultures with a positive growth index as determined by a continuous-reading automated blood culture system were examined by both conventional laboratory methods and FISH. For this purpose, oligonucleotide probes that allowed identification of approximately 95% of those pathogens typically associated with bacteremia were produced. The sensitivity and specificity of these probes were 100%. From all 115 blood cultures, microorganisms were grown after 1 day and identification to the family, genus, or species level was achieved after 1 to 3 days while 111 samples (96.5%) were similarly identified by FISH within 2.5 h. Staphylococci were identified in 62 of 62 samples, streptococci and enterococci were identified in 19 of 20 samples, gram-negative rods were identified in 28 of 30 samples, and fungi were identified in two of two samples. Thus, FISH is an appropriate method for identification of pathogens grown in blood cultures from septicemic patients. PMID:10655393

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

    PubMed Central

    Fredricks, David N.; Relman, David A.

    1998-01-01

    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 high-molecular-weight polyanion that is soluble in water but insoluble in alcohol. Accordingly, SPS tends to copurify with DNA. An extraction method was designed for purification of DNA from blood culture media and removal of SPS. Blood culture media containing human blood and spiked with Escherichia coli was subjected to an organic extraction procedure with benzyl alcohol, and removal of SPS was documented spectrophotometrically. Successful amplification of the extracted E. coli 16S rRNA gene was achieved by adding 5 μl of undiluted processed sample DNA to a 50-μl PCR mixture. When using other purification methods, the inhibitory effect of SPS could be overcome only by dilution of these samples. By our extraction technique, even uninoculated blood culture media were found to contain bacterial DNA when they were subjected to broad-range 16S rRNA gene consensus PCR. We conclude that the blood culture additive SPS is a potent inhibitor of PCR, is resistant to removal by traditional DNA purification methods, but can be removed by a benzyl alcohol extraction protocol that results in improved PCR performance. PMID:9738025

  2. High Mortality among Patients with Positive Blood Cultures at a Children's Hospital in Tbilisi, Georgia

    PubMed Central

    Schaffner, Jami; Chochua, Sopio; Kourbatova, Ekaterina V.; Barragan, Maribel; Wang, Yun F; Blumberg, Henry M; Rio, Carlos del; Walker, H. Kenneth; Leonard, Michael K.

    2010-01-01

    Background The etiology and outcomes of blood stream infections (BSI) among pediatric patients is not well described in resource-limited countries including Georgia. Methods Patients with positive blood cultures at the largest pediatric hospital in the country of Georgia were identified by review of medical and laboratory records for patients who had blood cultures obtained between 01/2004-06/2006. Results Of 1,693 blood cultures obtained during the study period, 338 (20%) were positive; 299 were included in our analysis. The median age was 14 days (range 2 days -14 years) and 178 (60%) were male; 53% of patients with a positive culture were admitted to Neonatal Intensive Care Unit (NICU). Gram-negative bacilli (GNB) were representing 165 (55%) of 299 cultures. Further speciation of 135 (82%) of 165 GNR was not possible because of lack of laboratory capacity. Overall mortality was 30% (90 of 299). Among the 90 children who died, 80 (89%) were neonates and 68 (76%) had BSI caused by Gram-negative organism. In multivariate analysis, independent risk factors for in-hospital mortality included age <30 days (OR=4.00, 95% CI 1.89-8.46) and having a positive blood culture for a Gram-negative BSI (OR=2.38, 95% CI 1.32-4.29). Conclusions A high mortality was seen among children, particularly neonates, with positive blood cultures at the largest pediatric hospital in Georgia. Because of limited laboratory capacity microbiological identification of common organisms known to cause BSI in children was not possible and susceptibility testing was not performed. Improving the infrastructure of diagnostic microbiology laboratories in resource limited countries is critical in order to improve patient care and clinical outcomes and from a public health standpoint to improve surveillance activities. PMID:19759489

  3. A Multicenter Evaluation of Blood Culture Practices, Contamination Rates, and the Distribution of Causative Bacteria

    PubMed Central

    Altindis, Mustafa; Koroglu, Mehmet; Demiray, Tayfur; Dal, Tuba; Ozdemir, Mehmet; Sengil, Ahmet Zeki; Atasoy, Ali Riza; Doğan, Metin; Cicek, Aysegul Copur; Ece, Gulfem; Kaya, Selcuk; Iraz, Meryem; Gultepe, Bilge Sumbul; Temiz, Hakan; Kandemir, Idris; Aksaray, Sebahat; Cetinkol, Yeliz; Sahin, Idris; Guducuoglu, Huseyin; Kilic, Abdullah; Kocoglu, Esra; Gulhan, Baris; Karabay, Oguz

    2016-01-01

    Background: The prognostic value of blood culture testing in the diagnosis of bacteremia is limited by contamination. Objectives: In this multicenter study, the aim was to evaluate the contamination rates of blood cultures as well as the parameters that affect the culture results. Materials and Methods: Sample collection practices and culture data obtained from 16 university/research hospitals were retrospectively evaluated. A total of 214,340 blood samples from 43,254 patients admitted to the centers in 2013 were included in this study. The blood culture results were evaluated based on the three phases of laboratory testing: the pre-analytic, the analytic, and the post-analytic phase. Results: Blood samples were obtained from the patients through either the peripheral venous route (64%) or an intravascular catheter (36%). Povidone-iodine (60%) or alcohol (40%) was applied to disinfect the skin. Of the 16 centers, 62.5% have no dedicated phlebotomy team, 68.7% employed a blood culture system, 86.7% conducted additional studies with pediatric bottles, and 43.7% with anaerobic bottles. One center maintained a blood culture quality control study. The average growth rate in the bottles of blood cultures during the defined period (1259 - 26,400/year) was 32.3%. Of the growing microorganisms, 67% were causative agents, while 33% were contaminants. The contamination rates of the centers ranged from 1% to 17%. The average growth time for the causative bacteria was 21.4 hours, while it was 36.3 hours for the contaminant bacteria. The most commonly isolated pathogens were Escherichia coli (22.45%) and coagulase-negative staphylococci (CoNS) (20.11%). Further, the most frequently identified contaminant bacteria were CoNS (44.04%). Conclusions: The high contamination rates were remarkable in this study. We suggest that the hospitals’ staff should be better trained in blood sample collection and processing. Sterile glove usage, alcohol usage for disinfection, the presence of

  4. [Confusion Over the Term "Contamination Rate" as It Pertains to Blood Cultures].

    PubMed

    Morii, Daiichi; Yokozawa, Takayuki; Ichinose, Naoki; Oda, Toshimi

    2016-05-01

    The blood culture contamination rate is often used to validate specimen-collection procedures. CUMITECH has set its optimal target to be 2% to 3%. However, the term "contamination rate" has been defined in many ways, limiting its generalizability. The definitions used in earlier studies can be divided into two categories; definitions based on clinical judgements, and those based on preset rules. According to each principle, the equation must be composed of a defined numerator and denominator. The problem with clinical definitions is that the decision is inevitably subjective, and the process is too cumbersome. Also, if the number of positive cultures is used as the denominator, the value would be equivalent to the positive predictive value, given that contamination is regarded as a "positive case." Thus, the value would not be useful for validating a procedure. On the other hand, when the preset algorithm was adopted, true infection would, to some degree, inevitably be classified as contamination. Also, if the algorithm adopted the number of blood culture sets as the denominator and contamination was defined as the identification of 1 or more specified organisms in only 1 of multiple sets of blood cultures, its theoretical maximum value would not be 100%. This is a problem because the value is a mixture of several numbers with different scales. In other words, whether the blood cultures are collected once, twice, or thrice or more a day would affect the result. The study cited by CUMITECH aimed to evaluate the equivalence between the clinical definition and the laboratory definition with preset rules, rather than to establish a benchmark for the contamination rate. It is undesirable for the number to be perceived as a benchmark. "A Guide to Blood Culture" (2013) by the Japanese Society for Clinical Microbiology introduced a calculation for the contamination rate, but the definition of the term "number of specimens" in the formula is ambiguous. In addition, the

  5. Blood culture contamination in hospitalized pediatric patients: a single institution experience

    PubMed Central

    Min, Hyewon; Park, Cheong Soo; Kim, Dong Soo

    2014-01-01

    Purpose Blood culture is the most important tool for detecting bacteremia in children with fever. However, blood culture contamination rates range from 0.6% to 6.0% in adults; rates for young children have been considered higher than these, although data are limited, especially in Korea. This study determined the contamination rate and risk factors in pediatric patients visiting the emergency room (ER) or being admitted to the ward. Methods We conducted a retrospective chart review of blood cultures obtained from children who visited Yonsei Severance Hospital, Korea between 2006 and 2010. Positive blood cultures were labeled as true bacteremia or contamination according to Centers for Disease Control and Prevention/National Healthcare Safety Network definitions for laboratory-confirmed bloodstream infection, after exclusion of cultures drawn from preexisting central lines only. Results Among 40,542 blood cultures, 610 were positive, of which 479 were contaminations and 131 were true bacteremia (overall contamination rate, 1.18%). The contamination rate in the ER was significantly higher than in the ward (1.32% vs. 0.66%, P<0.001). The rate was higher in younger children (2.07%, 0.94%, and 0.61% in children aged <1 year, 1-6 years, and >6 years, respectively). Conclusion Overall, contamination rates were higher in younger children than in older children, given the difficulty of performing blood sampling in younger children. The contamination rates from the ER were higher than those from the ward, not accounted for only by overcrowding and lack of experience among personnel collecting samples. Further study to investigate other factors affecting contamination should be required. PMID:24868215

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

    PubMed Central

    Pape, John; Wadlin, Jill; Nachamkin, Irving

    2006-01-01

    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. PMID:16825383

  7. High cell density media for Escherichia coli are generally designed for aerobic cultivations – consequences for large-scale bioprocesses and shake flask cultures

    PubMed Central

    Soini, Jaakko; Ukkonen, Kaisa; Neubauer, Peter

    2008-01-01

    Background For the cultivation of Escherichia coli in bioreactors trace element solutions are generally designed for optimal growth under aerobic conditions. They do normally not contain selenium and nickel. Molybdenum is only contained in few of them. These elements are part of the formate hydrogen lyase (FHL) complex which is induced under anaerobic conditions. As it is generally known that oxygen limitation appears in shake flask cultures and locally in large-scale bioreactors, function of the FHL complex may influence the process behaviour. Formate has been described to accumulate in large-scale cultures and may have toxic effects on E. coli. Although the anaerobic metabolism of E. coli is well studied, reference data which estimate the impact of the FHL complex on bioprocesses of E. coli with oxygen limitation have so far not been published, but are important for a better process understanding. Results Two sets of fed-batch cultures with conditions triggering oxygen limitation and formate accumulation were performed. Permanent oxygen limitation which is typical for shake flask cultures was caused in a bioreactor by reduction of the agitation rate. Transient oxygen limitation, which has been described to eventually occur in the feed-zone of large-scale bioreactors, was mimicked in a two-compartment scale-down bioreactor consisting of a stirred tank reactor and a plug flow reactor (PFR) with continuous glucose feeding into the PFR. In both models formate accumulated up to about 20 mM in the culture medium without addition of selenium, molybdenum and nickel. By addition of these trace elements the formate accumulation decreased below the level observed in well-mixed laboratory-scale cultures. Interestingly, addition of the extra trace elements caused accumulation of large amounts of lactate and reduced biomass yield in the simulator with permanent oxygen limitation, but not in the scale-down two-compartment bioreactor. Conclusion The accumulation of formate in

  8. Development and Evaluation of a Blood Culture PCR Assay for Rapid Detection of Salmonella Paratyphi A in Clinical Samples

    PubMed Central

    Zhou, Liqing; Jones, Claire; Gibani, Malick M.; Dobinson, Hazel; Thomaides-Brears, Helena; Shrestha, Sonu; Blohmke, Christoph J.; Darton, Thomas C.; Pollard, Andrew J.

    2016-01-01

    Background Enteric fever remains an important cause of morbidity in many low-income countries and Salmonella Paratyphi A has emerged as the aetiological agent in an increasing proportion of cases. Lack of adequate diagnostics hinders early diagnosis and prompt treatment of both typhoid and paratyphoid but development of assays to identify paratyphoid has been particularly neglected. Here we describe the development of a rapid and sensitive blood culture PCR method for detection of Salmonella Paratyphi A from blood, potentially allowing for appropriate diagnosis and antimicrobial treatment to be initiated on the same day. Methods Venous blood samples from volunteers experimentally challenged orally with Salmonella Paratyphi A, who subsequently developed paratyphoid, were taken on the day of diagnosis; 10 ml for quantitative blood culture and automated blood culture, and 5 ml for blood culture PCR. In the latter assay, bacteria were grown in tryptone soy broth containing 2.4% ox bile and micrococcal nuclease for 5 hours (37°C) before bacterial DNA was isolated for PCR detection targeting the fliC-a gene of Salmonella Paratyphi A. Results An optimized broth containing 2.4% ox bile and micrococcal nuclease, as well as a PCR test was developed for a blood culture PCR assay of Salmonella Paratyphi A. The volunteers diagnosed with paratyphoid had a median bacterial burden of 1 (range 0.1–6.9) CFU/ml blood. All the blood culture PCR positive cases where a positive bacterial growth was shown by quantitative blood culture had a bacterial burden of ≥ 0.3 CFU/ ml blood. The blood culture PCR assay identified an equal number of positive cases as automated blood culture at higher bacterial loads (≥0.3 CFU/ml blood), but utilized only half the volume of specimens. Conclusions The blood culture PCR method for detection of Salmonella Paratyphi A can be completed within 9 hours and offers the potential for same-day diagnosis of enteric fever. Using 5 ml blood, it exhibited a

  9. Blood

    MedlinePlus

    ... solid part of your blood contains red blood cells, white blood cells, and platelets. Red blood cells (RBC) deliver oxygen from your lungs to your tissues and organs. White blood cells (WBC) fight infection and are part of your ...

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

    PubMed

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

    2016-01-01

    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

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

    ERIC Educational Resources Information Center

    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…

  12. Evaluation of the necessity for routine terminal subculturing of blood cultures negative by radiometric methods.

    PubMed Central

    Beckwith, D G; Etowski, D C

    1982-01-01

    A retrospective review of 18,130 blood cultures performed with a BACTEC 225 indicated that terminal subculturing of bottles negative after 7 days of testing did not recover organisms which affected patient care or the length of patient hospitalization. PMID:7186906

  13. Rapid Intrinsic Fluorescence Method for Direct Identification of Pathogens in Blood Cultures

    PubMed Central

    Walsh, John D.; Hyman, Jay M.; Borzhemskaya, Larisa; Bowen, Ann; McKellar, Caroline; Ullery, Michael; Mathias, Erin; Ronsick, Christopher; Link, John; Wilson, Mark; Clay, Bradford; Robinson, Ron; Thorpe, Thurman; van Belkum, Alex; Dunne, W. Michael

    2013-01-01

    ABSTRACT A positive blood culture is a critical result that requires prompt identification of the causative agent. This article describes a simple method to identify microorganisms from positive blood culture broth within the time taken to perform a Gram stain (<20 min). The method is based on intrinsic fluorescence spectroscopy (IFS) of whole cells and required development of a selective lysis buffer, aqueous density cushion, optical microcentrifuge tube, and reference database. A total of 1,121 monomicrobial-positive broth samples from 751 strains were analyzed to build a database representing 37 of the most commonly encountered species in bloodstream infections or present as contaminants. A multistage algorithm correctly classified 99.6% of unknown samples to the Gram level, 99.3% to the family level, and 96.5% to the species level. There were no incorrect results given at the Gram or family classification levels, while 0.8% of results were discordant at the species level. In 8/9 incorrect species results, the misidentified isolate was assigned to a species of the same genus. This unique combination of selective lysis, density centrifugation, and IFS can rapidly identify the most common microbial species present in positive blood cultures. Faster identification of the etiologic agent may benefit the clinical management of sepsis. Further evaluation is now warranted to determine the performance of the method using clinical blood culture specimens. PMID:24255123

  14. Should Blood Cultures Be Performed for Patients with Acute Prostatitis? ▿

    PubMed Central

    Etienne, Manuel; Pestel-Caron, Martine; Chapuzet, Claire; Bourgeois, Ingrid; Chavanet, Pascal; Caron, François

    2010-01-01

    The diagnostic and prognostic values of blood cultures (BC) for 347 acute prostatitis inpatients were evaluated. BC were positive for 21% of patients and contributed to the microbiological diagnosis for 5%. Fever duration, length of hospitalization, use of an antibiotic combination, duration of antibiotic use, and urine bacterial titers increased when BC were positive. PMID:20237098

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

    PubMed

    Phoompoung, Pakpoom; Chayakulkeeree, Methee

    2016-06-01

    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

  16. Comparison of a homemade blood culture broth containing a papain digest of liver, with four commercially available media for the isolation of anaerobes from simulated paediatric blood cultures.

    PubMed Central

    Hunt, G H; Price, E H

    1982-01-01

    The recovery of small inocula of fastidious organisms, mainly non-sporing anaerobes, was studied in simulated paediatric blood culture experiments where only 1.5 ml of blood was added to each broth. A 25 ml homemade Queen Elizabeth Hospital medium (QEH medium) containing a papain digest of liver showed the best overall performance during the first four days of incubation; this medium was also satisfactory for maintenance of the majority of the organisms tested for longer than one week. LAB M Fastidious Anaerobe Broth (75 ml) with thymidine, also showed early isolation and satisfactory survival of most organisms. Difco Thiol broth, 50 ml with Liquoid, yielded early growth of the three strains of Bacteroides fragilis tested and maintained these organisms well; however, variable results were obtained with some other organisms in Difco Thiol media. Southern Group Brewer's thioglycollate broth (80 ml) gave the least satisfactory performance. PMID:6752208

  17. Evaluation of blood cultures in a children’s hospital located in Southeastern Anatolia

    PubMed Central

    Yiş, Reyhan

    2015-01-01

    Aim: Bloodstream infections in hospitalized patients are one of the most important causes of morbidity and mortality despite antimicrobial therapy. Early diagnosis and treatment of these infections is crucial. The aim of this study was to evaluate the distribution and antibiotic susceptibility of bacteria isolated from blood cultures in a children’s hospital in the Southeastern Anatolia during an 18-month period. Material and Methods: 7 040 blood cultures which were sent from hospitalized patients in Gaziantep Children’s Hospital between 01.07.2010 and 01.01.2012 were evaluated. Results: A total of 7 040 blood cultures were evaluated in this study. Microbial growth was detected in 2075 (29.47%) blood cultures. The most frequently isolated bacteria were coagulase-negative staphylococci (%45.97) which were followed by Salmonella spp. (%7.8). 12.12% of enterococcal isolates were resistant to glycopeptide antibiotics. The most frequently isolated gram negative bacterium was Salmonella spp. 15.43% of Salmonella spp. showed decreased susceptibility against quinolones. The ESBL positivity rate of E. coli and K. pneumoniae strains was found to be 35.08% and 57.14%, respectively. The imipenem resistance rate of P. aeruginosa was found to be 33.33%. The most common nonfermentative bacterium was S. maltophilia. Conclusions: The distribution of bacteria isolated from blood cultures and antibiotic resistance rates differ among different regions of Turkey. Different results obtained in our study may be related with regional tendencies to infections and patient population. Distribution of infectious agents and antibiotic resistance rates should be evaluated at regular intervals. This will lead to establishment of proper antibiotic usage policies in our country. PMID:26265894

  18. Direct Identification and Antimicrobial Susceptibility Testing of Bacteria From Positive Blood Culture Bottles by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry and the Vitek 2 System

    PubMed Central

    Jo, Sung Jin; Park, Kang Gyun; Han, Kyungja; Park, Dong Jin

    2016-01-01

    Background We evaluated the reliability and accuracy of the combined use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) bacterial identification and Vitek 2 antimicrobial susceptibility testing (AST) for bacteria from positive blood culture bottles. Methods Direct identification and AST were performed in parallel to the standard methods in monomicrobial positive blood culture bottles. In total, 254 isolates grown on aerobic and/or anaerobic bottles were identified with MALDI-TOF Vitek MS (bioMérieux, France), and 1,978 microorganism/antimicrobial agent combinations were assessed. For isolates from anaerobic bottles, an aliquot of the culture broth was centrifuged, washed, and filtered through a nylon mesh. For isolates from aerobic/pediatric bottles, a lysis step using 9.26% ammonium chloride solution and 2% saponin solution was included. Results The overall correct identification rate was 81.8% (208/254) and that for gram-positive/gram-negative isolates was 73.9%/92.6%, respectively, and it was 81.8%, 87.6%, and 57.9% for isolates from aerobic, anaerobic, and pediatric bottles, respectively. Identification was not possible in 45 cases, and most of these isolates were streptococci (N=14) and coagulase-negative staphylococci (N=11). Misidentification occurred only in one case. Compared with standard methods, direct AST showed 97.9% (1,936/1,978) agreement with very major error of 0.25%, major error of 0.05%, and minor error of 1.8%. Conclusions This simple and cost-effective sample preparation method gives reliable results for the direct identification and AST of bacteria. For the identification of streptococci and coagulase-negative staphylococci, the method should be further improved. PMID:26709258

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

    PubMed Central

    Chang, Chih-Jan; Wu, Chi-Jung; Hsu, Hsiang-Chin; Wu, Chiu-Hui; Shih, Fang-Ying; Wang, Shou-Wen; Wu, Yi-Hui; Chang, Chia-Ming; Tu, Yi-Fang; Chi, Chih-Hsien; Shih, Hsin-I

    2015-01-01

    Background 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. Methodology/Principal Findings 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. Conclusions/Significance 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. PMID:26448628

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

    PubMed Central

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

    2016-01-01

    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

  1. Effects of carbon sources on the enrichment of halophilic polyhydroxyalkanoate-storing mixed microbial culture in an aerobic dynamic feeding process

    PubMed Central

    Cui, You-Wei; Zhang, Hong-Yu; Lu, Peng-Fei; Peng, Yong-Zhen

    2016-01-01

    Microbial polyhydroxyalkanoate (PHA) production serves as a substitute for petroleum-based plastics. Enriching mixed microbial cultures (MMCs) with the capacity to store PHA is a key precursor for low-cost PHA production. This study investigated the impact of carbon types on enrichment outcomes. Three MMCs were separately fed by acetate sodium, glucose, and starch as an enriching carbon source, and were exposed to long-term aerobic dynamic feeding (ADF) periods. The PHA production capacity, kinetics and stoichiometry of the enrichments, the PHA composition, and the microbial diversity and community composition were explored to determine carbon and enrichment correlations. After 350-cycle enriching periods under feast-famine (F-F) regimes, the MMCs enriched by acetate sodium and glucose contained a maximum PHA content of 64.7% and 60.5% cell dry weight (CDW). The starch-enriched MMC only had 27.3% CDW of PHA. High-throughput sequencing revealed that non-PHA bacteria survived alongside PHA storing bacteria, even under severe F-F selective pressure. Genus of Pseudomonas and Stappia were the possible PHA accumulating bacteria in acetate-enriched MMC. Genus of Oceanicella, Piscicoccus and Vibrio were found as PHA accumulating bacteria in glucose-enriched MMC. Vibrio genus was the only PHA accumulating bacteria in starch-enriched MMC. The community diversity and composition were regulated by the substrate types. PMID:27485896

  2. Effects of carbon sources on the enrichment of halophilic polyhydroxyalkanoate-storing mixed microbial culture in an aerobic dynamic feeding process

    NASA Astrophysics Data System (ADS)

    Cui, You-Wei; Zhang, Hong-Yu; Lu, Peng-Fei; Peng, Yong-Zhen

    2016-08-01

    Microbial polyhydroxyalkanoate (PHA) production serves as a substitute for petroleum-based plastics. Enriching mixed microbial cultures (MMCs) with the capacity to store PHA is a key precursor for low-cost PHA production. This study investigated the impact of carbon types on enrichment outcomes. Three MMCs were separately fed by acetate sodium, glucose, and starch as an enriching carbon source, and were exposed to long-term aerobic dynamic feeding (ADF) periods. The PHA production capacity, kinetics and stoichiometry of the enrichments, the PHA composition, and the microbial diversity and community composition were explored to determine carbon and enrichment correlations. After 350-cycle enriching periods under feast-famine (F-F) regimes, the MMCs enriched by acetate sodium and glucose contained a maximum PHA content of 64.7% and 60.5% cell dry weight (CDW). The starch-enriched MMC only had 27.3% CDW of PHA. High-throughput sequencing revealed that non-PHA bacteria survived alongside PHA storing bacteria, even under severe F-F selective pressure. Genus of Pseudomonas and Stappia were the possible PHA accumulating bacteria in acetate-enriched MMC. Genus of Oceanicella, Piscicoccus and Vibrio were found as PHA accumulating bacteria in glucose-enriched MMC. Vibrio genus was the only PHA accumulating bacteria in starch-enriched MMC. The community diversity and composition were regulated by the substrate types.

  3. Effects of carbon sources on the enrichment of halophilic polyhydroxyalkanoate-storing mixed microbial culture in an aerobic dynamic feeding process.

    PubMed

    Cui, You-Wei; Zhang, Hong-Yu; Lu, Peng-Fei; Peng, Yong-Zhen

    2016-01-01

    Microbial polyhydroxyalkanoate (PHA) production serves as a substitute for petroleum-based plastics. Enriching mixed microbial cultures (MMCs) with the capacity to store PHA is a key precursor for low-cost PHA production. This study investigated the impact of carbon types on enrichment outcomes. Three MMCs were separately fed by acetate sodium, glucose, and starch as an enriching carbon source, and were exposed to long-term aerobic dynamic feeding (ADF) periods. The PHA production capacity, kinetics and stoichiometry of the enrichments, the PHA composition, and the microbial diversity and community composition were explored to determine carbon and enrichment correlations. After 350-cycle enriching periods under feast-famine (F-F) regimes, the MMCs enriched by acetate sodium and glucose contained a maximum PHA content of 64.7% and 60.5% cell dry weight (CDW). The starch-enriched MMC only had 27.3% CDW of PHA. High-throughput sequencing revealed that non-PHA bacteria survived alongside PHA storing bacteria, even under severe F-F selective pressure. Genus of Pseudomonas and Stappia were the possible PHA accumulating bacteria in acetate-enriched MMC. Genus of Oceanicella, Piscicoccus and Vibrio were found as PHA accumulating bacteria in glucose-enriched MMC. Vibrio genus was the only PHA accumulating bacteria in starch-enriched MMC. The community diversity and composition were regulated by the substrate types. PMID:27485896

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

    PubMed Central

    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.

    2016-01-01

    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 of Leptospira spp. 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

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

    PubMed

    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

    2016-04-01

    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 of Leptospira spp. 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

  6. Effects of transforming growth factor-beta on long-term human cord blood monocyte cultures

    SciTech Connect

    Orcel, P.; Bielakoff, J.; De Vernejoul, M.C. )

    1990-02-01

    Transforming growth factor-beta (TGF-beta) modulates growth and differentiation in many cell types and is abundant in bone matrix. We recently showed that human cord blood monocytes cultured in the presence of 1,25(OH)2D3 acquire some features of osteoclast precursors. Since TGF-beta has been shown to influence bone resorption in organ culture, we have studied the effect of TGF-beta (1-1,000 pg/ml) on cord blood monocyte cultures. These cells were cultured on plastic substrate during 3 weeks in the presence of 20% horse serum and 10(-9) M 1,25(OH)2D3. TGF-beta, from a concentration of 10 pg/ml in the culture medium, decreased in a dose dependent manner the formation of multinucleated cells. At a concentration of TGF-beta of 1 ng/ml, the multinucleated cells were reduced to 2.1% +/- 0.3%, compared to 19.3% +/- 1.5% in control cultures. TGF-beta inhibited in a dose-dependent manner the proliferation of cord blood monocytes as assessed by 3H-thymidine incorporation at 7 and 14 days of culture. The fusion index was also decreased by 3 weeks of treatment with TGF-beta. Indomethacin did not reverse the inhibitory effects of TGF-beta. The expression of the osteoclastic phenotype was assessed using two different antibodies: 23C6, a monoclonal antibody directed against the vitronectin receptor, which is highly expressed by osteoclasts but not by adult monocytes, and an antibody to HLA-DR, which is not present on osteoclast. TGF-beta decreased the expression of HLA-DR and increased in a dose-dependent manner the proportion of 23C6-labeled cells; these results suggest that TGF-beta could modulate a differentiation effect to the osteoclastic phenotype. However, when cord blood monocytes were cultured on devitalized rat calvariae prelabeled with 45Ca, TGF-beta did not induce any 45Ca release from bone cultured with monocytes.

  7. Diagnostic Accuracy of Procalcitonin for Predicting Blood Culture Results in Patients With Suspected Bloodstream Infection

    PubMed Central

    Oussalah, Abderrahim; Ferrand, Janina; Filhine-Tresarrieu, Pierre; Aissa, Nejla; Aimone-Gastin, Isabelle; Namour, Fares; Garcia, Matthieu; Lozniewski, Alain; Guéant, Jean-Louis

    2015-01-01

    Abstract Previous studies have suggested that procalcitonin is a reliable marker for predicting bacteremia. However, these studies have had relatively small sample sizes or focused on a single clinical entity. The primary endpoint of this study was to investigate the diagnostic accuracy of procalcitonin for predicting or excluding clinically relevant pathogen categories in patients with suspected bloodstream infections. The secondary endpoint was to look for organisms significantly associated with internationally validated procalcitonin intervals. We performed a cross-sectional study that included 35,343 consecutive patients who underwent concomitant procalcitonin assays and blood cultures for suspected bloodstream infections. Biochemical and microbiological data were systematically collected in an electronic database and extracted for purposes of this study. Depending on blood culture results, patients were classified into 1 of the 5 following groups: negative blood culture, Gram-positive bacteremia, Gram-negative bacteremia, fungi, and potential contaminants found in blood cultures (PCBCs). The highest procalcitonin concentration was observed in patients with blood cultures growing Gram-negative bacteria (median 2.2 ng/mL [IQR 0.6–12.2]), and the lowest procalcitonin concentration was observed in patients with negative blood cultures (median 0.3 ng/mL [IQR 0.1–1.1]). With optimal thresholds ranging from ≤0.4 to ≤0.75 ng/mL, procalcitonin had a high diagnostic accuracy for excluding all pathogen categories with the following negative predictive values: Gram-negative bacteria (98.9%) (including enterobacteria [99.2%], nonfermenting Gram-negative bacilli [99.7%], and anaerobic bacteria [99.9%]), Gram-positive bacteria (98.4%), and fungi (99.6%). A procalcitonin concentration ≥10 ng/mL was associated with a high risk of Gram-negative (odds ratio 5.98; 95% CI, 5.20–6.88) or Gram-positive (odds ratio 3.64; 95% CI, 3.11–4.26) bacteremia but

  8. Blood culture

    MedlinePlus

    ... symptoms of a serious infection, also known as sepsis . Symptoms of sepsis can include high fever, chills, rapid breathing and ... is bacteremia. This can be the result of sepsis. Sepsis is a medical emergency and you will ...

  9. Cost Effectiveness of the Antibiotic Removal Device for Processing Blood Cultures

    PubMed Central

    de Silva, Malkanthie I.; Qadri, S.M. Hussain; Hood, E.

    1986-01-01

    The antimicrobial removal device (ARD) blood culture system has been reported to increase the sensitivity of isolation of pathogenic microorganisms in bacteremic patients who are already on antibiotics. To determine the usefulness of this system to the clinician for the diagnosis of bacteremia and to determine the additional cost incurred by the use of the system, the microbiological results at two hospitals over a period of two years were compared. A total of 25,124 standard blood cultures (SBC) were performed with a positive culture rate of 10.7 percent. Of the 858 specimens processed by ARD alone, 68 (7.9 percent) were positive. There were a total of 2,657 specimens from 910 patients that were processed simultaneously using both systems. Both ARD and SBC were negative in 2,249 specimens, and 290 blood cultures from 107 patients grew the same organism using both systems. Thirty-one specimens from 12 patients grew pathogenic bacteria from ARD bottles; in each the SBC culture was negative. However, in 21 patients (44 specimens) bacteremia was detected only in SBC with negative cultures from ARD bottles. Thus, in the vast majority of the cases, SBC alone was sufficient to detect bacteremia, even in the patient with recent or concomitant antibiotic therapy. The total processing cost was calculated for the cases in which SBC and ARD were performed simultaneously and was found to be $6,588 for SBC and $15,005 for ARD. The comparative cost per bacteremic patient detected by the two methods was $46.40 for SBC and $555.75 for ARD. PMID:3097333

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

    PubMed

    Desai, Sunil G

    2015-11-10

    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

  11. Isolation and characterization of aerobic culturable arsenic-resistant bacteria from surfacewater and groundwater of Rautahat District, Nepal.

    PubMed

    Shakya, S; Pradhan, B; Smith, L; Shrestha, J; Tuladhar, S

    2012-03-01

    Arsenic (As) contamination of groundwater is a serious Environmental Health Management issue of drinking water sources especially in Terai region of Nepal. Many studies have reported that due to natural abundance of arsenic in the environment, various bacteria have developed different resistance mechanisms for arsenic compound. In this study, the culturable arsenic-resistant bacteria indigenous to surfacewater as well as groundwater from Rautahat District of Nepal were randomly isolated by standard plate count method on the basis of viable growth on plate count agar amended with arsenate ranging from 0, 0.5, 10, 40, 80 to 160 milligram per liter (mg/l). With respect to the morphological and biochemical tests, nine morphologically distinct potent arsenate tolerant bacteria showed relatedness with Micrococcus varians, Micrococcus roseus, Micrococcus luteus, Pseudomonas maltophilia, Pseudomonas sp., Vibrio parahaemolyticus, Bacillus cereus, Bacillus smithii 1 and Bacillus smithii 2. The isolates were capable of tolerating more than 1000 mg/l of arsenate and 749 mg/l of arsenite. Likewise, bioaccumulation capability was highest with M. roseus (85.61%) and the least with B. smithii (47.88%) indicating the potential of the organisms in arsenic resistance and most probably in bioremediation. PMID:21868146

  12. Blood Cultures for Persistent Fever in Neutropenic Pediatric Patients Are of Low Diagnostic Yield.

    PubMed

    Neemann, Kari; Yonts, Alexandra B; Qiu, Fang; Simonsen, Kari; Lowas, Stefanie; Freifeld, Alison

    2016-06-01

    The incidence of bacteremia at the onset of pediatric febrile neutropenia (FN) at 2 academically linked institutions was 9.84%, and subsequent blood cultures performed for children with persistent FN yielded an incidence of 4.21%. Until the risk factors for new-onset bacteremia in patients being treated for FN can be identified and diagnostic methods can be improved, compliance with national guidelines is recommended. PMID:27199474

  13. Impact of Hourly Emergency Department Patient Volume on Blood Culture Contamination and Diagnostic Yield

    PubMed Central

    Halverson, Schuyler; Malani, Preeti N.; Newton, Duane W.; Habicht, Andrea; Vander Have, Kenneth

    2013-01-01

    Emergency departments (EDs) are an important diagnostic site for outpatients with potentially serious infections. EDs frequently experience high patient volumes, and crowding has been shown to negatively impact the delivery of early care for serious infections, such as pneumonia. Here, we hypothesized that other important factors in the early care of infectious diseases, the rate of blood culture contamination and the accurate detection of pathogens, would be sensitive to ED operational stress, as proper collection requires fastidious attention to technique and timing. We related all blood samples collected over 1 year and their rates of recovery of likely contaminants and pathogens to the number of patients being cared for in the ED at the time of sample collection. Likely pathogens and contaminants were classified through combined microbiological and manual chart review criteria. Zero-inflated Poisson regression was used to relate crowding to culture results. Blood samples were obtained from 7,586 patients over 82,521 adult and pediatric patient visits. The unadjusted rates of recovering a likely pathogen or a likely contaminant were 8.0% and 3.7%, respectively. Periods of increased crowding (3rd and 4th quartiles of hourly occupancy) were significantly associated (P < 0.01) with increased rates of contamination (relative risk, 1.23 compared to the least busy quartile). Collecting samples for culture during busy times was also associated with a reduced likelihood of recovering a likely pathogen (relative risk, 0.93 compared to the least busy quartile). ED crowding was associated with degraded performance of blood cultures, both increasing the rate of contamination and decreasing the diagnostic yield. PMID:23515549

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

    PubMed

    Schukur, Lina; Geering, Barbara; Fussenegger, Martin

    2016-03-01

    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

  15. Skin antiseptics in venous puncture-site disinfection for prevention of blood culture contamination: systematic review with meta-analysis.

    PubMed

    Caldeira, D; David, C; Sampaio, C

    2011-03-01

    Blood cultures drawn by venous puncture are common clinical procedures for the detection of bacteraemia. Blood culture contamination (BCC) can lead to clinical misinterpretation and unnecessary expenses. We aimed to systematically review randomised controlled trials (RCTs) with skin antiseptics for prevention of contamination in venous-puncture drawn blood cultures. We conducted database search using CENTRAL (Cochrane Library issue April 2010), MEDLINE, EMBASE and mRCT, in June 2010. All RCTs testing skin antiseptics in venous-puncture drawn blood cultures were retrieved. Relative risk (RR) of the BCC outcome was analysed by random effects method using confidence interval (CI) of 95%. Studies were assessed by one review author and checked by another. Six studies were identified. Single-trial comparisons showed that alcoholic iodine tincture was better than non-alcoholic povidone-iodine, and isopropyl/acetone/povidone-iodine showed superiority against isopropyl/povidone-iodine. Meta-analysis demonstrated that alcoholic chlorhexidine was better than non-alcoholic povidone-iodine (RR: 0.33; 95% CI: 0.24-0.46) in 4757 blood cultures from two trials. Alcoholic solutions were better than non-alcoholic products (0.53; 0.31-0.90) in 21,300 blood cultures from four studies. Two trials with 13,418 blood cultures showed that iodine tincture was not superior to povidone-iodine in BCC prevention (0.79; 0.54-1.18). Alcoholic iodine was not different from non-alcoholic iodine (0.79; 0.53-1.17). Comparison of chlorhexidine vs iodine compounds was not conclusive. Alcohol alone was not inferior to iodinated products for prevention of contamination in venous-puncture drawn blood cultures. The association of alcohol and povidone-iodine did not seem to be useful. Alcoholic chlorhexidine solutions reduced blood culture false positives compared with aqueous povidone-iodine. PMID:21194791

  16. A bovine mammary endothelial/epithelial cell culture model of the blood/milk barrier.

    PubMed Central

    Guidry, A J; O'Brien, C N; Douglass, L W

    1998-01-01

    The complex nature of the mammary gland has hampered in-depth studies of the relationship of the circulatory system to cells lining the teat ducts and alveoli of the gland. This study reports an in vitro model of endothelial and epithelial cells separated by a subcellular matrix that simulates the blood milk barrier of the bovine mammary gland. Dual chamber culture dishes with a porous membrane separating the upper and lower chamber were used. Endothelial and epithelial cells were cultured on opposite sides of the porous membrane. A collagen and fibroblast subcellular matrix, separating the 2 cell layers, simulated the in vivo interstitial tissue. Changes in surface binding of anti-bodies to polymorphonuclear neutrophils (PMN) following their migration from the upper to the lower chamber simulated the passage of PMN from blood to milk. Changes in the binding of antibodies to PMN agreed with results observed following the migration of PMN from blood to milk in vivo. This gives credence to the model's potential value for studies where more direct observation of the blood/milk barrier is required. The model will be further tested for its usefulness as an assay for determining: 1) antibiotic diffusion from milk to blood and from blood to milk, 2) cytotoxicity of prophylactic and therapeutic mammary infusion products, 3) factors affecting bacterial adhesion and penetration of mammary epithelial tissue, 4) effectiveness of antibodies present in lacteal secretions in preventing bacterial adhesion, and 5) the feasibility of gene constructs to induce synthesis and secretion of mastitis-preventing compounds and prophylactic and therapeutic compounds for treatment of human disorders. PMID:9553710

  17. Blood

    MedlinePlus

    ... fight infection and are part of your body's defense system. Platelets help blood to clot when you have a cut or wound. Bone marrow, the spongy material inside your bones, makes new blood cells. Blood cells ...

  18. Isolation of Corynebacterium tuscaniae sp. nov. from Blood Cultures of a Patient with Endocarditis

    PubMed Central

    Riegel, Philippe; Creti, Roberta; Mattei, Romano; Nieri, Alfredo; von Hunolstein, Christina

    2006-01-01

    A strain of an unknown coryneform bacterium was repeatedly isolated in pure culture from the blood of a patient affected by endocarditis. Comparative 16S rRNA gene sequence analysis revealed that this isolate represented a new subline within the genus Corynebacterium. This new taxon can be identified by the presence of corynomycolic acids and its enzymatic activities and fermentation of sugars. Acid production from glucose and maltose, pyrazinamidase and alkaline phoshatase activities, and hippurate hydrolysis were the most characteristic phenotypic features of the bacterium. On the basis of both phenotypic and phylogenetic evidence, it is proposed that this isolate be classified as a novel species, Corynebacterium tuscaniae sp. nov. The type strain, ISS-5309, has been deposited in the American Type Culture Collection (ATCC BAA-1141) and in the Culture Collection of the University of Göteborg (CCUG 51321). PMID:16455875

  19. Effects of Culture Conditions and Tuberculin Source on Interferon-gamma production in Whole Blood Cultures from Mycobacterium bovis Infected Cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The BOVIGAM® interferon (IFN) - gamma assay constitutes an ante-mortem, in vitro laboratory-based tuberculosis test and is used complementary to the tuberculin skin test. The assay is performed in two stages: firstly, whole blood is cultured with antigens stimulating blood leucocytes to produce IFN...

  20. Effects of Culture Conditions and Tuberculin Source on Interferon-gamma Production in Whole Blood Cultures from Mycobacterium bovis Infected Cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The BOVIGAM® interferon (IFN) - gamma assay constitutes an ante-mortem, in vitro laboratory-based tuberculosis test and is used complementary to the tuberculin skin test. The assay is performed in two stages: firstly, whole blood is cultured with antigens stimulating blood leucocytes to produce IFN-...

  1. [Stimulation of cell cultures recovery after cryopreservation by the cattle cord blood FRACTION (below 5 kDa) or Actovegin].

    PubMed

    2013-01-01

    The capacities of the cattle cord blood low-molecular fraction (below 5 kDa) and Actovegin (the vealer blood fraction (below 5 kDa)) for recovering functions of cell cultures after cryopreservation compared. Their influence proliferation of the flozen-thawed cell cultures, certain stages of their growth, cell attachment, rate of cell spreading, and mitotic regiment has been studied. Both the cord blood low-molecular fraction and Actovegin were shown to stimulate growth of the cell cultures after cryopreservation more efficiently at the concentration of 224 μg/ml. However, despite the stimulating effect discovered, their application did not bring proliferative indices on the 1st passage after cryopreservation to the values of the native culture. The effects of the cord blood low-molecular fraction and Actovegin on the human fibroblast culture were identical by the following parameters: cell attachment, rates of cell spreading and proliferation. In culture BHK-21 clone 13/04 the efficiency of Actovegin was low, while the cord blood low-molecular fraction has a conspicuous stimulating effect on its adhesion and proliferation. The investigations carried out can serve as a basis for the development of regenerative media containing the cattle cord blood low-molecular fraction (below 5 kDa) or Actovegin as active components at the concentration of 224 μg/ml with the purpose of fast recovery of culture prolifetative properties after cryopreservation. PMID:25508566

  2. [Stimulation of cell cultures recovery after cryopreservation by the cattle cord blood FRACTION (below 5 kDa) or Actovegin].

    PubMed

    Gulevskiĭ, A K; Trifonova, A V; Lavrik, A A

    2013-01-01

    The capacities of the cattle cord blood low-molecular fraction (below 5 kDa) and Actovegin (the vealer blood fraction (below 5 kDa)) for recovering functions of cell cultures after cryopreservation compared. Their influence proliferation of the flozen-thawed cell cultures, certain stages of their growth, cell attachment, rate of cell spreading, and mitotic regiment has been studied. Both the cord blood low-molecular fraction and Actovegin were shown to stimulate growth of the cell cultures after cryopreservation more efficiently at the concentration of 224 μg/ml. However, despite the stimulating effect discovered, their application did not bring proliferative indices on the 1st passage after cryopreservation to the values of the native culture. The effects of the cord blood low-molecular fraction and Actovegin on the human fibroblast culture were identical by the following parameters: cell attachment, rates of cell spreading and proliferation. In culture BHK-21 clone 13/04 the efficiency of Actovegin was low, while the cord blood low-molecular fraction has a conspicuous stimulating effect on its adhesion and proliferation. The investigations carried out can serve as a basis for the development of regenerative media containing the cattle cord blood low-molecular fraction (below 5 kDa) or Actovegin as active components at the concentration of 224 μg/ml with the purpose of fast recovery of culture prolifetative properties after cryopreservation. PMID:25470939

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

    PubMed

    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

    2016-05-01

    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

  4. Cytogenetic and oxidative alterations after exposure of cultured human whole blood cells to lithium metaborate dehydrate.

    PubMed

    Çelikezen, Fatih Çağlar; Toğar, Başak; Özgeriş, Fatma Betül; İzgi, Mehmet Sait; Türkez, Hasan

    2016-08-01

    Boron compounds have an ability of supporting antioxidant properties in human and animal tissues. Lithium metaborate dihydrate (LiBO2·2H2O; LMD) is commonly used in nonlinear optic materials, cellular phones and pagers. But, there are limited data on the genotoxic and antioxidant effects of LMD in cultured human whole blood cells. The aim of this study was to evaluate for the genotoxicity and antioxidant/oxidant activity of LMD on human whole blood lymphocytes (n = 5). LMD was applied at various concentrations (0-1,280 µg/ml) to cultured blood samples. Antioxidant/oxidant activity was evaluated by measuring the total oxidant status (TOS) and total antioxidant capacity levels. Micronuclei and chromosomal aberration tests were used in genotoxicity studies. Our results clearly revealed that all tested concentrations of LMD were found to be non-genotoxic when compared to that of the control group. In addition, LMD exhibited antioxidant activities at low concentrations. In addition the TOS levels were not changed at all concentrations of LMD. Consequently, our results clearly demonstrated that LMD is non-genotoxic and it has an important antioxidant potential in vitro. PMID:25680697

  5. Revisiting the Diego Blood Group System in Amerindians: Evidence for Gene-Culture Comigration.

    PubMed

    Bégat, Christophe; Bailly, Pascal; Chiaroni, Jacques; Mazières, Stéphane

    2015-01-01

    Six decades ago the DI*A allele of the Diego blood group system was instrumental in proving Native American populations originated from Siberia. Since then, it has received scant attention. The present study was undertaken to reappraise distribution of the DI*A allele in 144 Native American populations based on current knowledge. Using analysis of variance tests, frequency distribution was studied according to geographical, environmental, and cultural parameters. Frequencies were highest in Amazonian populations. In contrast, DI*A was undetectable in subarctic, Fuegian, Panamanian, Chaco and Yanomama populations. Closer study revealed a correlation that this unequal distribution was correlated with language, suggesting that linguistic divergence was a driving force in the expansion of DI*A among Native Americans. The absence of DI*A in circumpolar Eskimo-Aleut and Na-Dene speakers was consistent with a late migratory event confined to North America. Distribution of DI*A in subtropical areas indicated that gene and culture exchanges were more intense within than between ecozones. Bolstering the utility of classical genetic markers in biological anthropology, the present study of the expansion of Diego blood group genetic polymorphism in Native Americans shows strong evidence of gene-culture comigration. PMID:26148209

  6. Revisiting the Diego Blood Group System in Amerindians: Evidence for Gene-Culture Comigration

    PubMed Central

    Bégat, Christophe; Bailly, Pascal; Chiaroni, Jacques; Mazières, Stéphane

    2015-01-01

    Six decades ago the DI*A allele of the Diego blood group system was instrumental in proving Native American populations originated from Siberia. Since then, it has received scant attention. The present study was undertaken to reappraise distribution of the DI*A allele in 144 Native American populations based on current knowledge. Using analysis of variance tests, frequency distribution was studied according to geographical, environmental, and cultural parameters. Frequencies were highest in Amazonian populations. In contrast, DI*A was undetectable in subarctic, Fuegian, Panamanian, Chaco and Yanomama populations. Closer study revealed a correlation that this unequal distribution was correlated with language, suggesting that linguistic divergence was a driving force in the expansion of DI*A among Native Americans. The absence of DI*A in circumpolar Eskimo-Aleut and Na-Dene speakers was consistent with a late migratory event confined to North America. Distribution of DI*A in subtropical areas indicated that gene and culture exchanges were more intense within than between ecozones. Bolstering the utility of classical genetic markers in biological anthropology, the present study of the expansion of Diego blood group genetic polymorphism in Native Americans shows strong evidence of gene-culture comigration. PMID:26148209

  7. The Optimization of Molecular Detection of Clinical Isolates of Brucella in Blood Cultures by eryD Transcriptase Gene for Confirmation of Culture-Negative Samples

    PubMed Central

    Tabibnejad, Mahsa; Alikhani, Mohammad Yousef; Arjomandzadegan, Mohammad; Hashemi, Seyed Hamid; Naseri, Zahra

    2016-01-01

    Background Brucellosis is a zoonosis disease which is widespread across the world. Objectives The aim of the present study is the evaluation of culture-negative blood samples. Materials and Methods A total of 100 patients with suspected brucellosis were included in this experimental study and given positive serological tests. Diagnosis was performed on patients with clinical symptoms of the disease, followed by the detection of a titer that was equal to or more than 1:160 (in endemic areas) by the standard tube agglutination method. Blood samples were cultured by a BACTEC 9050 system, and subsequently by Brucella agar. At the same time, DNA from all blood samples was extracted by Qiagen Kit Company (Qia Amp Mini Kit). A molecular assay of blood samples was carried out by detection of eryD transcriptase and bcsp 31 genes in specific double PCR reactions. The specificity of the primers was evaluated by DNA from pure and approved Brucella colonies found in the blood samples, by DNA from other bacteria, and by ordinary PCR. DNA extraction from the pure colonies was carried out by both Qiagen Kit and Chelex 100 methods; the two were compared. Results 39 cases (39%) had positive results when tested by the BACTEC system, and 61 cases (61%) became negative. 23 culture-positive blood samples were randomly selected for PCR reactions; all showed 491 bp for the eryD gene and 223 bp for the bcsp 31 gene. Interestingly, out of 14 culture-negative blood samples, 13 cases showed positive bonds in PCR. The specificity of the PCR method was equal to 100%. DNA extraction from pure cultures was done by both Chelex 100 and Qiagen Kit; these showed the same results for all samples. Conclusions The results prove that the presented double PCR method could be used to detect positive cases from culture-negative blood samples. The Chelex 100 method is simpler and safer than the use of Qiagen Kit for DNA extraction. PMID:27330831

  8. Burkholderia gladioli infection isolated from the blood cultures of newborns in the neonatal intensive care unit.

    PubMed

    Zhou, F; Ning, H; Chen, F; Wu, W; Chen, A; Zhang, J

    2015-08-01

    Burkholderia gladioli was described as a plant pathogen, and it is a rare cause of infection in humans that is primarily associated with human pulmonary infections, such as chronic granulomatous disease and cystic fibrosis. The neonatal respiratory system is not fully developed and cannot expel bacterial aerosol properly. A total of 2,676 newborns in the neonatal intensive care unit were retrospectively analysed in Putian City, Fujian Province, China, from 2011 to 2014. All of the blood samples were tested for C-reactive protein (CRP), procalcitonin (PCT) and white blood cell (WBC). B. gladioli infections were determined and analysed using a blood culture system. Antibiotic susceptibility testing was performed using the K-B method. Of the 2,676 participants, 87 (3.25 %) had a positive B. gladioli blood culture that occurred >72 h after birth, including a premature group (54.0 %, asphyxia [vs. 9.20 %], fever [vs. 13.80 %], pneumonia [vs. 6.90 %] and hyperbilirubinaemia [vs. 8.05 %]) and newborns with necrotising enterocolitis (NEC) (vs. 5.75 %). The mean ± standard deviation (SD) of the CRP level was 12.31 ± 0.26 mg/L and that of the PCT level was 1.53 ± 0.21 ng/ml in the 87 B. gladioli-infected newborns. Most of the B. gladioli isolates were sensitive to many antimicrobial agents and did not lead to serious consequences. All of the B. gladioli-infected newborns were unhealthy, especially the premature infants. B. gladioli might be a causative bacteraemia agent in neonates, it appears to have pathogenic potential in newborns and its sensitivity to antibiotics may be a beneficial factor. PMID:25926303

  9. Blood-culture-proven neonatal septicaemia: a review of 36 cases.

    PubMed

    Misallati, A; el-Bargathy, S; Shembesh, N

    2000-01-01

    The cases of 36 newborns seen in the neonatal unit of Al-Fatah Children's Hospital in Benghazi, Libyan Arab Jamahiriya, with blood-culture-proven septicaemia were reviewed to determine clinical profile and outcome. There were 22 males and 14 females. Of these, 12 infants were premature with a gestational age of < 37 weeks and 24 were full term (gestational age > 37 weeks). At diagnosis, 11 cases were under 4 days of age. The most common symptoms were lethargy and feeding intolerance. Klebsiella was the most common etiological microorganism. Bacterial isolates were resistant to ampicillin and gentamicin but sensitive to cefotaxime. Of the 36 infants, 12 died (fatality rate = 33%). PMID:11556040

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

    PubMed

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

    2016-01-01

    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

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

    PubMed

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

    2014-01-01

    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. PMID:24713904

  12. Proportional-Integral-Derivative (PID) Control of Secreted Factors for Blood Stem Cell Culture

    PubMed Central

    Caldwell, Julia; Wang, Weijia; Zandstra, Peter W.

    2015-01-01

    Clinical use of umbilical cord blood has typically been limited by the need to expand hematopoietic stem and progenitor cells (HSPC) ex vivo. This expansion is challenging due to the accumulation of secreted signaling factors in the culture that have a negative regulatory effect on HSPC output. Strategies for global regulation of these factors through dilution have been developed, but do not accommodate the dynamic nature or inherent variability of hematopoietic cell culture. We have developed a mathematical model to simulate the impact of feedback control on in vitro hematopoiesis, and used it to design a proportional-integral-derivative (PID) control algorithm. This algorithm was implemented with a fed-batch bioreactor to regulate the concentrations of secreted factors. Controlling the concentration of a key target factor, TGF-β1, through dilution limited the negative effect it had on HSPCs, and allowed global control of other similarly-produced inhibitory endogenous factors. The PID control algorithm effectively maintained the target soluble factor at the target concentration. We show that feedback controlled dilution is predicted to be a more cost effective dilution strategy compared to other open-loop strategies, and can enhance HSPC expansion in short term culture. This study demonstrates the utility of secreted factor process control strategies to optimize stem cell culture systems, and motivates the development of multi-analyte protein sensors to automate the manufacturing of cell therapies. PMID:26348930

  13. 3D cultured immortalized human hepatocytes useful to develop drugs for blood-borne HCV

    SciTech Connect

    Aly, Hussein Hassan; Shimotohno, Kunitada; Hijikata, Makoto

    2009-02-06

    Due to the high polymorphism of natural hepatitis C virus (HCV) variants, existing recombinant HCV replication models have failed to be effective in developing effective anti-HCV agents. In the current study, we describe an in vitro system that supports the infection and replication of natural HCV from patient blood using an immortalized primary human hepatocyte cell line cultured in a three-dimensional (3D) culture system. Comparison of the gene expression profile of cells cultured in the 3D system to those cultured in the existing 2D system demonstrated an up-regulation of several genes activated by peroxisome proliferator-activated receptor alpha (PPAR{alpha}) signaling. Furthermore, using PPAR{alpha} agonists and antagonists, we also analyzed the effect of PPAR{alpha} signaling on the modulation of HCV replication using this system. The 3D in vitro system described in this study provides significant insight into the search for novel anti-HCV strategies that are specific to various strains of HCV.

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

    PubMed

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

    2016-05-01

    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

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

    PubMed

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

    2016-07-01

    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

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

    PubMed Central

    Mascart, G; Bertrand, F; Mascart, P

    1983-01-01

    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 acridine orange yields more positive results than Gram staining and is also simpler. PMID:6188764

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

    PubMed Central

    Chihara, Shingo; Hayden, Mary K.; Minogue-Corbett, Eileen; Singh, Kamaljit

    2009-01-01

    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. PMID:20016679

  18. Prevalence of extended-spectrum beta-lactamase-producing Enterobacteriaceae isolated from blood cultures in Africa.

    PubMed

    Sangare, S A; Maiga, A I; Guindo, I; Maiga, A; Camara, N; Savadogo, S; Diallo, S; Bougoudogo, F; Armand-Lefevre, L; Andremont, A; Maiga, I I

    2015-09-01

    Extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae have been isolated from many regions of the world. Epidemiological studies are being conducted in Europe, North America, and Asia. No study has however been conducted in Africa to determine the prevalence and distribution of ESBLs on the continent. This literature review aimed at describing the prevalence of ESBL-producing Enterobacteriaceae isolated from blood cultures, as well as the ESBL genes involved at the international level. Our focus was mainly on Africa. We conducted a literature review on PubMed. Articles related to our study field and published between 1996 and 2014 were reviewed and entirely read for most of them, while we only focused on the abstracts of some other articles. Relevant articles to our study were then carefully reviewed and included in the review. The prevalence of ESBL-producing Enterobacteriaceae differs from one country to another. The results of our literature review however indicate that class A ESBLs prevail over the other types. We took into consideration articles focusing on various types of samples to assess the prevalence of ESBL-producing Enterobacteriaceae, but information on isolates from blood cultures is limited. The worldwide prevalence of ESBL-producing Enterobacteriaceae has increased over time. Evidence of ESBL-producing Enterobacteriaceae can be found in all regions of the world. Studies conducted in Africa mainly focused on the Northern and Eastern parts of the continent, while only rare studies were carried out in the rest of the continent. PMID:26433872

  19. Teaching Aerobic Fitness Concepts.

    ERIC Educational Resources Information Center

    Sander, Allan N.; Ratliffe, Tom

    2002-01-01

    Discusses how to teach aerobic fitness concepts to elementary students. Some of the K-2 activities include location, size, and purpose of the heart and lungs; the exercise pulse; respiration rate; and activities to measure aerobic endurance. Some of the 3-6 activities include: definition of aerobic endurance; heart disease risk factors;…

  20. Time to Detection with BacT/Alert FA Plus Compared to BacT/Alert FA Blood Culture Media.

    PubMed

    Nutman, A; Fisher Even-Tsur, S; Shapiro, G; Braun, T; Schwartz, D; Carmeli, Y

    2016-09-01

    Rapid identification of the causative pathogen in patients with bacteremia allows adjustment of antibiotic therapy and improves patient outcomes. We compared in vitro and real-life time to detection (TTD) of two blood culture media, BacT/Alert FA (FA) and BacT/Alert FA Plus (FA Plus), for the nine most common species of bacterial pathogens recovered from blood samples. Experimental data from simulated cultures was compared with microbiology records of TTD for both culture media with growth of the species of interest in clinical blood cultures. In the experimental conditions, median TTD was 3.8 hours (23.9 %) shorter using FA Plus media. The magnitude of reduction differed between species. Similarly, in real life data, FA Plus had shorter TTD than FA media; however, the difference between culture media was smaller, and median TTD was only 1 hour (8.5 %) less. We found shorter TTD with BacT/Alert FA Plus culture media, both experimentally and in real-life conditions and unrelated to antibiotic neutralization, highlighting the importance of appropriate blood culture media selection. PMID:27272123

  1. Nanobacteria from blood: the smallest culturable autonomously replicating agent on Earth

    NASA Astrophysics Data System (ADS)

    Kajander, E. Olavi; Kuronen, Ilpo; Akerman, Kari K.; Pelttari, Alpo; Ciftcioglu, Neva

    1997-07-01

    Nanobacteria are the first mineral forming bacteria isolated from blood and blood products. They are coccoid cell-walled organisms with a size of 0.08 - 0.5 micrometers in EM, occure in clusters, produce a biofilm containing carbonate or hydroxyl apatite, and are highly resistant to heat, gamma-irradiation and antibiotics. Their growth rate is about one hundredth that of ordinary bacteria and they divide via several mechanisms. Taq polymerase was able to use their nontraditional nucleic acid as a template. 16S rRNA gene sequence results positioned them into the alpha-2 subgroup of Proteobacteria. Nanobacteria are smallest cell-walled bacteria since they can pass through 0.07 micrometers pores. In low-serum cultures, they form even smaller elementary particles or tubular units. How can blood be infected with such slow growing, heat and radio-resistant bacteria? The answer may lie in their phylogeny: alpha-2 subgroup has organisms from soil exposed to radiation and heat, that can penetrate into eukaryotic cells. Nanobacteria grow so slowly that they require a niche `cleaned' with heat, radiation or immunodefence. For survival they cloak themselves in apatite, a normal constituent of mammalian body. This may link nanobacteria to nannobacteria discovered from sedimentary rocks by Dr. Folk. Both have similar size, size variation, clustering and mineral deposits. They may resemble the probable ancient bacterial fossils in the Martian meteorite ALH84001.

  2. NanoFlares for the detection, isolation, and culture of live tumor cells from human blood.

    PubMed

    Halo, Tiffany L; McMahon, Kaylin M; Angeloni, Nicholas L; Xu, Yilin; Wang, Wei; Chinen, Alyssa B; Malin, Dmitry; Strekalova, Elena; Cryns, Vincent L; Cheng, Chonghui; Mirkin, Chad A; Thaxton, C Shad

    2014-12-01

    Metastasis portends a poor prognosis for cancer patients. Primary tumor cells disseminate through the bloodstream before the appearance of detectable metastatic lesions. The analysis of cancer cells in blood—so-called circulating tumor cells (CTCs)—may provide unprecedented opportunities for metastatic risk assessment and investigation. NanoFlares are nanoconstructs that enable live-cell detection of intracellular mRNA. NanoFlares, when coupled with flow cytometry, can be used to fluorescently detect genetic markers of CTCs in the context of whole blood. They allow one to detect as few as 100 live cancer cells per mL of blood and subsequently culture those cells. This technique can also be used to detect CTCs in a murine model of metastatic breast cancer. As such, NanoFlares provide, to our knowledge, the first genetic-based approach for detecting, isolating, and characterizing live cancer cells from blood and may provide new opportunities for cancer diagnosis, prognosis, and personalized therapy. PMID:25404304

  3. Isolation, Culture, and Characterization of Human Umbilical Cord Blood-Derived Mesenchymal Stromal Cells.

    PubMed

    Bieback, Karen; Netsch, Philipp

    2016-01-01

    Umbilical cord blood (CB) is considered one of the youngest available sources of adult stem cells. Besides hematopoietic stem cells, CB has been shown to contain endothelial progenitor cells as well as mesenchymal stromal/stem cells (MSC). To isolate MSC from cord blood, CB is collected into a sterile bag containing the anticoagulant citrate-phosphate-dextrose (CPD). The CB is then processed by density-gradient centrifugation to obtain mononuclear cells (MNC). These are cultured until the outgrowth of fibroblastoid cell colonies appears. After reaching a subconfluent stage, cells are harvested, expanded, and characterized as cord blood mesenchymal stromal cells (CB-MSC) according to standard criteria: plastic adherence, fibroblast morphology, CFU-f assay, proliferation potential, immune phenotype, and differentiation potential.Apparently, the frequency of MSC in CB is extremely low. Thus, not every CB unit will provide adequate MSC isolation yields. Different strategies have been proposed aiming to optimize the isolation success by selecting CB units of optimal quality. It is commonly agreed on that a high CB volume, a high cellular content, and a short time frame between birth and MSC isolation are criteria that will enhance the MSC isolation success.The procedures in this chapter are standardized protocols that were established and optimized in the authors' research laboratory; however, various modifications of the protocols are possible. PMID:27236676

  4. Effects of an artificial gravity countermeasure on orthostatic tolerance, blood volumes and aerobic power after short-term bed rest (BR-AG1).

    PubMed

    Linnarsson, Dag; Hughson, Richard L; Fraser, Katelyn S; Clément, Gilles; Karlsson, Lars L; Mulder, Edwin; Paloski, William H; Rittweger, Jörn; Wuyts, Floris L; Zange, Jochen

    2015-01-01

    Exposure to artificial gravity (AG) in a short-arm centrifuge has potential benefits for maintaining human performance during long-term space missions. Eleven subjects were investigated during three campaigns of 5 days head-down bed rest: 1) bed rest without countermeasures (control), 2) bed rest and 30 min of AG (AG1) daily, and 3) bed rest and six periods of 5 min AG (AG2) daily. During centrifugation, the supine subjects were exposed to AG in the head-to-feet direction with 1 G at the center of mass. Subjects participated in the three campaigns in random order. The cardiovascular effects of bed rest and countermeasures were determined from changes in tolerance to a head-up tilt test with superimposed lower body negative pressure (HUT), from changes in plasma volume (PV) and from changes in maximum aerobic power (V̇o2 peak) during upright work on a cycle ergometer. Complete data sets were obtained in eight subjects. After bed rest, HUT tolerance times were 36, 64, and 78% of pre-bed rest baseline during control, AG1 and AG2, respectively, with a significant difference between AG2 and control. PV and V̇o2 peak decreased to 85 and 95% of pre-bed rest baseline, respectively, with no differences between the treatments. It was concluded that the AG2 countermeasure should be further investigated during future long-term bed rest studies, especially as it was better tolerated than AG1. The superior effect of AG2 on orthostatic tolerance could not be related to concomitant changes in PV or aerobic power. PMID:25342708

  5. Antibiotic utilization improvement with the Nanosphere Verigene Gram-Positive Blood Culture assay.

    PubMed

    Beal, Stacy G; Thomas, Cody; Dhiman, Neelam; Nguyen, Daniel; Qin, Huanying; Hawkins, Jennifer M; Dekmezian, Mhair; Benavides, Raul

    2015-04-01

    New technologies offer rapid identification of organisms and antimicrobial resistance markers in blood cultures several hours faster than conventional methods. We sought to determine whether implementation of the Verigene® Gram-Positive Blood Culture (BC-GP) assay paired with a well-defined results reporting algorithm would lead to earlier deescalation of empiric therapy for inpatients with methicillin-sensitive Staphylococcus aureus (MSSA) and vancomycin-resistant Enterococcus (VRE) bacteremia. The algorithm design focused on lessening the demand for pharmacist time by using electronic communications where possible. Our study compared inpatients with MSSA and VRE bacteremia from the time period before (pre-BC-GP) and after (post-BC-GP) implementation of the assay on June 25, 2013. The time from blood draw to identification and susceptibility results was decreased by 36.4 hours (P < 0.001) in the post-BC-GP group. The mean time from collection to the first dose of optimal antibiotics was reduced in the post-BC-GP group by 18.9 hours (P = 0.004) overall, with a 20.6-hour reduction (P = 0.009) for patients with MSSA and a 20.7-hour reduction (P = 0.077) for patients with VRE. Additionally, the percent of patients on empiric therapy who were placed on optimal antibiotics at any time after the Gram stain result was available increased from 64% (45/70) pre-BC-GP to 80% (43/54) post-BC-GP. The BC-GP led to an increased rate of deescalation of empiric antibiotics and a reduction in the time to optimal antibiotics for patients with MSSA and VRE bacteremia. PMID:25829639

  6. Controlled clinical laboratory comparison of BACTEC plus aerobic/F resin medium with BacT/Alert aerobic FAN medium for detection of bacteremia and fungemia.

    PubMed Central

    Jorgensen, J H; Mirrett, S; McDonald, L C; Murray, P R; Weinstein, M P; Fune, J; Trippy, C W; Masterson, M; Reller, L B

    1997-01-01

    Blood specimens collected from adult patients with suspected sepsis in four medical centers were inoculated into BACTEC Plus/F and BacT/Alert FAN aerobic culture bottles. Both bottles of 7,401 bottle pairs contained the prescribed blood volume of 8 to 12 ml. Bottles were incubated in their respective instruments for a standard 7-day protocol or until the instruments signaled that they were positive. A total of 720 isolates that were judged to represent true infections were recovered from 338 patients; 451 isolates were recovered from both bottles, 143 were recovered from only the Plus/F bottle, and 126 were recovered from only the FAN bottle (P was not significant). Although more Histoplasma capsulatum isolates were recovered from Plus/F bottles (P < 0.005), there were no other statistically significant differences in recovery rates of individual species or groups of organisms between the two systems. Of 329 monomicrobic patient septic episodes, 244 episodes were detected by both blood culture systems, 40 were detected only by the BACTEC system, and 45 were detected only by the BacT/Alert system (P was not significant). There was no significant difference between the two systems in the detection of septic episodes among patients receiving antibiotic therapy at the time of blood cultures. Of the cultures found to be positive within the first 72 h of incubation, detection was on average earlier by the BACTEC system (16.9 h) than by the BacT/Alert system (18.7 h). Larger differences in average time to detection were seen with streptococci (10.7 h by the BACTEC system and 17.9 h by the BacT/Alert system) and yeasts (an average of 29.4 h by the BacT/Alert system versus 37.2 h by the BACTEC system). With the exception of the differences noted above, BACTEC Plus/F aerobic resin and BacT/Alert aerobic FAN blood culture bottles were comparable in their abilities to recover aerobic and facultative organisms. PMID:8968880

  7. Comparison of BACTEC PLUS Blood Culture Media to BacT/Alert FA Blood Culture Media for Detection of Bacterial Pathogens in Samples Containing Therapeutic Levels of Antibiotics▿

    PubMed Central

    Flayhart, Diane; Borek, Anita P.; Wakefield, Teresa; Dick, James; Carroll, Karen C.

    2007-01-01

    Blood culture bottles with antimicrobial removal systems are recommended for patients who develop fever while on antibiotics. This study compared the ability of Becton Dickinson (Sparks, MD) BACTEC PLUS bottles and bioMerieux (Durham, NC) BacT/Alert FA bottles to effectively remove vancomycin, cefoxitin, ceftriaxone, cefepime, piperacillin-tazobactam, ampicillin, oxacillin, gentamicin, and a combination of gentamicin/penicillin, thus allowing bacterial pathogens to grow. Each bottle was spiked with 10 ml of human blood, antibiotic, and strains of organisms susceptible to the antibiotic evaluated. The organisms used were type strains and clinical isolates of Staphylococcus aureus (methicillin susceptible and resistant), Streptococcus pneumoniae, a viridans streptococcus, Enterococcus faecalis, Enterococcus faecium, Streptococcus agalactiae, Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Testing was completed in triplicate, using 10 to 100 CFU/ml of organisms with various concentrations of each antibiotic. Two rounds of testing were completed per antibiotic/organism combination. Bottles were mixed and loaded onto their respective instruments as per the manufacturer's instructions. Antimicrobial removal was evaluated on the basis of time to detection of organism growth, for up to 5 days of incubation. Overall, the BacT/Alert FA system recovered 25.1% of strains from test bottles and 96.9% of strains from growth control bottles (no antibiotic added), and the BACTEC PLUS system recovered 95.1% of strains from test bottles and 100% of strains from growth control bottles. Both systems performed well in the detection of Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa in the presence of gentamicin. In the presence of ceftriaxone, neither system was able to recover Streptococcus pneumoniae. The ability to remove vancomycin and cefoxitin was also determined by measuring antibiotic levels remaining in bottles after 1 h of incubation

  8. Comparison of BACTEC PLUS blood culture media to BacT/Alert FA blood culture media for detection of bacterial pathogens in samples containing therapeutic levels of antibiotics.

    PubMed

    Flayhart, Diane; Borek, Anita P; Wakefield, Teresa; Dick, James; Carroll, Karen C

    2007-03-01

    Blood culture bottles with antimicrobial removal systems are recommended for patients who develop fever while on antibiotics. This study compared the ability of Becton Dickinson (Sparks, MD) BACTEC PLUS bottles and bioMerieux (Durham, NC) BacT/Alert FA bottles to effectively remove vancomycin, cefoxitin, ceftriaxone, cefepime, piperacillin-tazobactam, ampicillin, oxacillin, gentamicin, and a combination of gentamicin/penicillin, thus allowing bacterial pathogens to grow. Each bottle was spiked with 10 ml of human blood, antibiotic, and strains of organisms susceptible to the antibiotic evaluated. The organisms used were type strains and clinical isolates of Staphylococcus aureus (methicillin susceptible and resistant), Streptococcus pneumoniae, a viridans streptococcus, Enterococcus faecalis, Enterococcus faecium, Streptococcus agalactiae, Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Testing was completed in triplicate, using 10 to 100 CFU/ml of organisms with various concentrations of each antibiotic. Two rounds of testing were completed per antibiotic/organism combination. Bottles were mixed and loaded onto their respective instruments as per the manufacturer's instructions. Antimicrobial removal was evaluated on the basis of time to detection of organism growth, for up to 5 days of incubation. Overall, the BacT/Alert FA system recovered 25.1% of strains from test bottles and 96.9% of strains from growth control bottles (no antibiotic added), and the BACTEC PLUS system recovered 95.1% of strains from test bottles and 100% of strains from growth control bottles. Both systems performed well in the detection of Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa in the presence of gentamicin. In the presence of ceftriaxone, neither system was able to recover Streptococcus pneumoniae. The ability to remove vancomycin and cefoxitin was also determined by measuring antibiotic levels remaining in bottles after 1 h of incubation

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

    NASA Astrophysics Data System (ADS)

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

    2005-05-01

    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.

  10. Vi-specific latex agglutination for early and rapid detection of Salmonella serotype typhi in blood cultures.

    PubMed

    Jesudason, M V; Sridharan, G; Mukundan, S; John, T J

    1994-02-01

    Latex particles coated with rabbit antisera against Salmonella serotype typhi (S. typhi) Vi and O (STO) antigens were used in slide agglutination tests for the rapid identification of S. typhi in blood culture broths as soon as Gram-negative bacilli (GNB) were detected in them. Among 231 consecutive blood cultures showing GNB tested for Vi, and a subset of 163 tested for STO, by latex agglutination (LA), 125 and 32, respectively, were positive. The GNB in 127 blood cultures were confirmed by conventional methods as S. typhi, 125 (98.4%) of which had been identified by the Vi LA test. In the subset of 163, 81 grew S. typhi, of which only 32 (39.5%) had been identified by the STO LA tests. Thus, the sensitivity of the Vi and STO LA tests was 98.4% and 39.5%, respectively, whereas the specificity was 100% for both tests. Of the S. typhi isolates, 38 (30.4%) were detected by the Vi LA test on day 2 and 73 (58.4%) on day 3, day 1 being the date of inoculation of the blood culture broths. Thus, the Vi LA test is suitable for the early and rapid confirmation of S. typhi in blood culture. PMID:7520382

  11. Bacterial Wound Culture

    MedlinePlus

    ... Home Visit Global Sites Search Help? Bacterial Wound Culture Share this page: Was this page helpful? Also known as: Aerobic Wound Culture; Anaerobic Wound Culture Formal name: Culture, wound Related ...

  12. Investigation of oxidative phosphorylation in continuous cultures. A non-equilibrium thermodynamic approach to energy transduction for Escherichia coli in aerobic condition

    NASA Astrophysics Data System (ADS)

    Ghafuri, Mohazabeh; Nosrati, Mohsen; Hosseinkhani, Saman

    2015-03-01

    Adenosine triphosphate (ATP) production in living cells is very important. Different researches have shown that in terms of mathematical modeling, the domain of these investigations is essentially restricted. Recently the thermodynamic models have been suggested for calculation of the efficiency of oxidative phosphorylation process and rate of energy loss in animal cells using chemiosmotic theory and non-equilibrium thermodynamics equations. In our previous work, we developed a mathematical model for mitochondria of animal cells. In this research, according to similarities between oxidative phosphorylation process in microorganisms and animal cells, Golfar's model was developed to predict the non-equilibrium thermodynamic behavior of the oxidative phosphorylation process for bacteria in aerobic condition. With this model the rate of energy loss, P/O ratio, and efficiency of oxidative phosphorylation were calculated for Escherichia coli in aerobic condition. The results then were compared with experimental data given by other authors. The thermodynamic model had an acceptable agreement with the experimental data.

  13. Comparative in vitro activity of ceftaroline, ceftaroline-avibactam, and other antimicrobial agents against aerobic and anaerobic bacteria cultured from infected diabetic foot wounds.

    PubMed

    Goldstein, Ellie J C; Citron, Diane M; Merriam, C Vreni; Tyrrell, Kerin L

    2013-07-01

    Foot infections are the most common infectious complication of diabetes. Moderate to severe diabetic foot infections (DFI) are typically polymicrobial with both aerobic and anaerobic organisms. The role of MRSA in these wounds has become an increasing concern. To determine if the addition of avibactam, a novel non-beta-lactam beta-lactamase inhibitor, to ceftaroline would be more active than ceftaroline alone, we tested 316 aerobic pathogens and 154 anaerobic recovered from patients with moderate to severe DFI, and compared ceftaroline with and without avibactam to other agents. Testing on aerobes was done by broth microdilution and by agar dilution for anaerobes, according to CLSI M11-A8, and M7-A8 standards. Ceftaroline-avibactam MIC90 for all Staphylococcus spp. including MRSA was 0.5 μg/mL, and for enterococci was 1 μg/mL. The MIC90s for enteric Gram-negative rods was 0.125 μg/mL. The addition of avibactam to ceftaroline reduced the ceftaroline MICs for 2 strains of resistant Enterobacter spp. and for 1 strain of Morganella. Against anaerobic Gram-positive cocci ceftaroline-avibactam had an MIC90 0.125 μg/mL and for clostridia 1 μg/mL. Avibactam improved ceftaroline's MIC90s for Bacteroides fragilis from >32 to 2 μg/mL and for Prevotella spp. from >32 to 1 μg/mL. Ceftaroline alone demonstrates excellent in vitro activity against most of the aerobes found in moderate to severe DFI. The addition of avibactam provides an increased spectrum of activity including the beta-lactamase producing Prevotella, Bacteroides fragilis and ceftaroline resistant gram-negative enteric organisms. PMID:23623385

  14. [Native valve Aspergillus fumigatus endocarditis with blood culture positive and negative for galactomannan antigen. Case report and literature review].

    PubMed

    Pemán, Javier; Ortiz, Rebeca; Osseyran, Faisa; Pérez-Bellés, Carmen; Crespo, Marisa; Chirivella, Melitina; Frasquet, Juan; Quesada, Anastasio; Cantón, Emilia; Gobernado, Miguel

    2007-06-01

    Native valve endocarditis caused by Aspergillus spp. is an uncommon disease with a high mortality rate. Generally, Aspergillus is isolated from affected valve in post-mortem or biopsy specimens. However, its isolation from blood cultures is exceedingly rare. We report a case of fungal endocarditis in a native mitral valve with the isolation of Aspergillus fumigatus both in valve vegetation and in blood culture bottles. The patient underwent valve replacement and antifungal treatment with voriconazole and caspofungin, but he died on post-operative day 45 with disseminated aspergillosis confirmed by necropsy. Paradoxically, galactomannan antigen detection in serum was negative. This is the third case of Aspergillus endocarditis with positive blood culture reported in the literature. PMID:17604438

  15. In vitro culture and differentiation of osteoblasts from human umbilical cord blood.

    PubMed

    Toai, Tran Cong; Thao, Huynh Duy; Thao, Nguyen Phuong; Gargiulo, Ciro; Ngoc, Phan Kim; Van, Pham Hung; Strong, D Michael

    2010-08-01

    It is well accepted that human umbilical cord blood (UCB) is a source of mesenchymal stem cells (MSCs) which are able to differentiate into different cell phenotypes such as osteoblasts, chondrocytes, adipocytes, myocytes, cardiomyocytes and neurons. The aim of this study was to isolate MSCs from human UCB to determine their osteogenic potential by using different kinds of osteogenic medium. Eventually, only those MSCs cultured in osteogenic media enriched with vitamin D(2) and FGF9, were positive for osteocalcin by RT-PCR. All these cells were positive for alizarin red, alkaline phosphatase and Von Kossa. The results obtained from RT-PCR have confirmed that osteogenesis is complete by expression of the osteocalcin marker. In conclusion, vitamin D(2), at least in vitro, may replace vitamin D(3) as an osteogenic stimulator factor for MSC differentiation. PMID:19565355

  16. Importance of blood cultures to aid the diagnosis of Lemierre's syndrome.

    PubMed

    Nejat, Maryam; Werno, Anja

    2015-05-15

    This is a case report of Lemierre's syndrome, a septic thrombophlebitis of the internal jugular vein (IJV) usually preceded by pharyngitis and bacteraemia with an anaerobic organism. Fusobacterium necrophorum is ananaerobic Gram-negative bacillus and is the most common organism reported to cause Lemierre's syndrome which usually occurs one to three weeks post pharyngitis or oropharyngeal surgery. A 21-year-old patient presented with signs of sepsis and a history of sore throat, fever, and tender cervical lymph nodes. Blood cultures grew F. necrophorum and Computed Tomography (CT) showed a filling defect in the left retromandibular vein and thrombosis in the left internal jugular vein (IJV) consistent with Lemierre's syndrome. This is an uncommon condition which normally occurs in young individuals and diagnosis is often delayed. PMID:26117393

  17. Dimethyl Sulfoxide Enhances Effectiveness of Skin Antiseptics and Reduces Contamination Rates of Blood Cultures

    PubMed Central

    LaSala, Paul R.; Han, Xiang-Yang; Rolston, Kenneth V.; Kontoyiannis, Dimitrios P.

    2012-01-01

    Effective skin antisepsis is of central importance in the prevention of wound infections, colonization of medical devices, and nosocomial transmission of microorganisms. Current antiseptics have a suboptimal efficacy resulting in substantial infectious morbidity, mortality, and increased health care costs. Here, we introduce an in vitro method for antiseptic testing and a novel alcohol-based antiseptic containing 4 to 5% of the polar aprotic solvent dimethyl sulfoxide (DMSO). The DMSO-containing antiseptic resulted in a 1- to 2-log enhanced killing of Staphylococcus epidermidis and other microbes in vitro compared to the same antiseptic without DMSO. In a prospective clinical validation, blood culture contamination rates were reduced from 3.04% for 70% isopropanol–1% iodine (control antiseptic) to 1.04% for 70% isopropanol–1% iodine–5% DMSO (P < 0.01). Our results predict that improved skin antisepsis is possible using new formulations of antiseptics containing strongly polarized but nonionizing (polar aprotic) solvents. PMID:22378911

  18. Identification and susceptibility testing of Staphylococcus aureus by direct inoculation from positive BACTEC blood culture bottles.

    PubMed

    Diederen, B M W; Zieltjens, M; Wetten, H; Buiting, A G M

    2006-01-01

    This study explored the possibility of combining direct inoculation of tube coagulase and DNase tests, and the VITEK2 system, from BACTEC blood culture bottles in order to achieve rapid identification and susceptibility testing of Staphylococcus aureus. All isolates were identified correctly as S. aureus or coagulase-negative staphylococci (CNS). Antimicrobial susceptibility testing with the VITEK2 system gave 99.6% correct category agreement, with 0.1% very major errors and 0.3% minor errors among S. aureus isolates, and 97.4% correct category agreement, with 0.9% very major errors and 1.7% minor errors among CNS isolates. The results suggested that direct identification and susceptibility testing is sufficiently accurate for immediate reporting. PMID:16460552

  19. Autologous red blood cells potentiate antibody synthesis by unfractionated human mononuclear cell cultures.

    PubMed

    Rugeles, M T; La Via, M; Goust, J M; Kilpatrick, J M; Hyman, B; Virella, G

    1987-08-01

    We have tried to determine the most favourable conditions for the in vitro induction of specific antibody (Ab) responses to tetanus toxoid (TT) and keyhole limpet haemocyanin (KLH). Human peripheral blood mononuclear cells (PBMNC) were obtained from normal volunteers and stimulated with PWM, TT, KLH, and mixtures of PWM and antigens in the presence or absence of autologous red blood cells (RBC) (1:50 ratio of PBMNC/RBC). The cultures were harvested on day 11; immunoglobulins were determined immunonephelometrically and Ab levels by ELISA with human antibodies used for calibration. While anti-TT responses were easy to induce with PBMNC from recently boosted individuals, the production of anti-TT from PBMNC obtained from non-recently boosted individuals was only possible when PBMNC were stimulated with TT and PWM in the presence of autologous RBC. Similarly, anti-KLH responses were easier to induce with PBMNC from an immune donor; maximal response was observed after stimulation with PWM + KLH in the presence of autologous RBC. Stimulation of primary anti-KLH responses with PBMNC from non-immune donors was only successful when the cells were stimulated with KLH + PWM in the presence of autologous RBC. The potentiation of human B-cell responses with autologous RBC can be abrogated by pretreatment of PBMNC with anti-CD2 antibodies and is associated with increased expression of IL-2 receptors and increased production of gamma interferon (IFN-gamma). However, addition of IFN-gamma in different doses and at different times to PWM-stimulated PBMNC cultures was not as effective as addition of RBC in enhancing the production of immunoglobulin and antibody. PMID:3114872

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

    PubMed

    Aoki, Yosuke

    2010-05-01

    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

  1. EFFECTS OF PRE-CULTURE HOLDING TIME AND TEMPERATURE ON INTERFERON-GAMMA RESPONSES IN WHOLE BLOOD CULTURES FROM MYCOBACTERIUM BOVIS-INFECTED CATTLE

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Bovigam™ assay is approved for use within the United States as a complimentary test for tuberculosis. Prior to whole blood culture and the ensuing ELISA to detect interferon- (IFN) ', samples are subjected to various holding time / temperature combinations due, in part, to practical constraints ...

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

    PubMed

    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

    2012-02-24

    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

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

    PubMed

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

    2016-01-01

    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

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

    PubMed Central

    Weinstein, M P; Reller, L B; Mirrett, S; Stratton, C W; Reimer, L G; Wang, W L

    1986-01-01

    A commercially available agar-slide blood culture bottle (Septi-Chek; Roche Diagnostics, Div. Hoffman-La Roche, Inc., Nutley, N.J.) was compared with the radiometric blood culture system (BACTEC; Johnston Laboratories, Inc., Towson, Md.) in 8,544 paired blood cultures from adult patients. The systems were inoculated with equal volumes (10 ml) of blood. Overall, there was no statistically significant difference between the two systems in the recovery of clinically important microorganisms, but significantly more members of the family Enterobacteriaceae other than Escherichia coli were detected by the agar-slide system (P less than 0.005). The agar-slide system detected more fungi, and the BACTEC detected more anaerobic bacteria; however, small numbers of recovered organisms precluded statistical significance. When microorganisms grew in both systems, their presence was detected one or more days earlier in the BACTEC (P less than 0.001). More contaminants grew in the agar-slide system (P less than 0.001). Both systems performed well, and either system should provide high yield and prompt detection of positive blood cultures in patients with bacteremia and fungemia if used in an optimal way as recommended by the respective manufacturers. PMID:3517047

  5. How to Optimize the Use of Blood Cultures for the Diagnosis of Bloodstream Infections? A State-of-the Art

    PubMed Central

    Lamy, Brigitte; Dargère, Sylvie; Arendrup, Maiken C.; Parienti, Jean-Jacques; Tattevin, Pierre

    2016-01-01

    Bloodstream infection (BSI) is a major cause of death in developed countries and the detection of microorganisms is essential in managing patients. Despite major progress has been made to improve identification of microorganisms, blood culture (BC) remains the gold standard and the first line tool for detecting BSIs. Consensus guidelines are available to ensure optimal BSI procedures, but BC practices often deviate from the recommendations. This review provides an update on clinical and technical issues related to blood collection and to BC performance, with a special focus on the blood sample strategy to optimize the sensitivity and specificity of BCs. PMID:27242721

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

    PubMed Central

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

    2010-01-01

    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

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

    PubMed

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

    2015-05-01

    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

  8. Performance Evaluation of the Verigene Gram-Positive and Gram-Negative Blood Culture Test for Direct Identification of Bacteria and Their Resistance Determinants from Positive Blood Cultures in Hong Kong

    PubMed Central

    Siu, Gilman K. H.; Chen, Jonathan H. K.; Ng, T. K.; Lee, Rodney A.; Fung, Kitty S. C.; To, Sabrina W. C.; Wong, Barry K. C.; Cheung, Sherman; Wong, Ivan W. F.; Tam, Marble M. P.; Lee, Swing S. W.; Yam, W. C.

    2015-01-01

    Background A multicenter study was conducted to evaluate the diagnostic performance and the time to identifcation of the Verigene Blood Culture Test, the BC-GP and BC-GN assays, to identify both Gram-positive and Gram-negative bacteria and their drug resistance determinants directly from positive blood cultures collected in Hong Kong. Methods and Results A total of 364 blood cultures were prospectively collected from four public hospitals, in which 114 and 250 cultures yielded Gram-positive and Gram-negative bacteria, and were tested with the BC-GP and BC-GN assay respectively. The overall identification agreement for Gram-positive and Gram-negative bacteria were 89.6% and 90.5% in monomicrobial cultures and 62.5% and 53.6% in polymicrobial cultures, respectively. The sensitivities for most genus/species achieved at least 80% except Enterococcus spp. (60%), K.oxytoca (0%), K.pneumoniae (69.2%), whereas the specificities for all targets ranged from 98.9% to 100%. Of note, 50% (7/14) cultures containing K.pneumoniae that were missed by the BC-GN assay were subsequently identified as K.variicola. Approximately 5.5% (20/364) cultures contained non-target organisms, of which Aeromonas spp. accounted for 25% and are of particular concern. For drug resistance determination, the Verigene test showed 100% sensitivity for identification of MRSA, VRE and carbapenem resistant Acinetobacter, and 84.4% for ESBL-producing Enterobacteriaceae based on the positive detection of mecA, vanA, blaOXA and blaCTXM respectively. Conclusion Overall, the Verigene test provided acceptable accuracy for identification of bacteria and resistance markers with a range of turnaround time 40.5 to 99.2 h faster than conventional methods in our region. PMID:26431434

  9. Blood cell oxidative stress precedes hemolysis in whole blood-liver slice co-cultures of rat, dog, and human tissues

    SciTech Connect

    Vickers, Alison E.M.; Sinclair, John R.; Fisher, Robyn L.; Morris, Stephen R.; Way, William

    2010-05-01

    A novel in vitro model to investigate time-dependent and concentration-dependent responses in blood cells and hemolytic events is studied for rat, dog, and human tissues. Whole blood is co-cultured with a precision-cut liver slice. Methimazole (MMI) was selected as a reference compound, since metabolism of its imidazole thione moiety is linked with hematologic disorders and hepatotoxicity. An oxidative stress response occurred in all three species, marked by a decline in blood GSH levels by 24 h that progressed, and preceded hemolysis, which occurred at high MMI concentrations in the presence of a liver slice with rat (>= 1000 muM at 48 h) and human tissues (>= 1000 muM at 48 h, >= 750 muM at 72 h) but not dog. Human blood-only cultures exhibited a decline of GSH levels but minimal to no hemolysis. The up-regulation of liver genes for heme degradation (Hmox1 and Prdx1), iron cellular transport (Slc40a1), and GSH synthesis and utilization (mGST1 and Gclc) were early markers of the oxidative stress response. The up-regulation of the Kupffer cell lectin Lgals3 gene expression indicated a response to damaged red blood cells, and Hp (haptoglobin) up-regulation is indicative of increased hemoglobin uptake. Up-regulation of liver IL-6 and IL-8 gene expression suggested an activation of an inflammatory response by liver endothelial cells. In summary, MMI exposure led to an oxidative stress response in blood cells, and an up-regulation of liver genes involved with oxidative stress and heme homeostasis, which was clearly separate and preceded frank hemolysis.

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

    PubMed

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

    2014-05-01

    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 blood culture growing S. aureus during 2002 through 2003 (pre intervention n = 58) or during 2008 through 2009 (post intervention n = 100) were included. Medical records were evaluated regarding clinical data and outcome. Blood culture isolates were characterized by antibiotic susceptibility testing (AST) and S. aureus protein A (spa) gene typing. The annual incidence of S. aureus bacteremia (SAB) increased from 28 per 100,000 inhabitants at the pre intervention period to 45 per 100,000 at the post intervention period (p = 0.046). During post intervention, the SAB incidence was significantly higher in men (p = 0.009). The mortality rate during hospital stay was 14 % during pre intervention and 18 % during post intervention (p = 0.47). The most common spa types were t012 and t084. The Surviving Sepsis Campaign resulted in an increased number of detected cases of SAB. The mortality rate was the same before and after the intervention, and no spa type correlated to certain clinical manifestations or mortality. PMID:24249284

  11. RAPID IDENTIFICATION OF CANDIDA ALBICANS DIRECTLY FROM YEAST POSITIVE BLOOD CULTURE BOTTLES BY FLUORESCENCE IN SITU HYBRIDIZATION USING PNA PROBES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

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

    PubMed

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

    2011-11-01

    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

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

    PubMed Central

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

    2011-01-01

    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

  14. Evaluation of canine and feline leishmaniasis by the association of blood culture, immunofluorescent antibody test and polymerase chain reaction

    PubMed Central

    2014-01-01

    Background This study aimed to evaluate the occurrence of Leishmania spp. in dogs and cats from Botucatu, São Paulo state, and Campo Grande, Mato Grosso do Sul state, Brazil, by the association of three diagnostic tests: blood culture in liver infusion tryptose medium, immunofluorescent antibody test and polymerase chain reaction. Fifty blood samples of dogs and cats from the Center for Zoonosis Control in Campo Grande, an area endemic for canine visceral leishmaniasis, were collected randomly, as well as canine and feline blood samples from the Municipal Kennel and Animal Protection Association in Botucatu, currently considered a transmission-free, non-endemic area. Results Of the 50 dog blood cultures from Botucatu, three (6%) were positive and of the 50 cats, two (4%) were positive. In Campo Grande, 29 dog blood cultures (58%) were positive and all (100%) cats negative by this test. Polymerase chain reaction detected Leishmania spp. in 100% of dog and cat samples from Botucatu but found all the cats from Campo Grande to be negative. On the other hand, 36 dogs from Campo Grande were positive (72%) by the same technique. Immunofluorescent antibody test in Botucatu found 100% of dogs and cats non-reactive, while in Campo Grande, it detected positivity in 32 dogs (64%) and 15 cats (30%). Conclusions The results show the importance of not only continuous epidemiological surveillance in areas not endemic for leishmaniasis, but also research for accurate diagnosis of this zoonosis. PMID:24565284

  15. Evaluation of Real-time PCR and Pyrosequencing for Screening Incubating Blood Culture Bottles from Adults with Suspected Bloodstream Infection

    PubMed Central

    McCann, Chase D.; Moore, Miranda S.; May, Larissa S.; McCarroll, Matthew; Jordan, Jeanne A.

    2015-01-01

    Several molecular platforms can identify bacteria associated with bloodstream infections, but require positive culture bottles as starting material. Here we describe results of screening 1140 blood cultures at 8 hours post-inoculation, from 918 eligible adults being evaluated for bloodstream infection. DNA was extracted and analyzed by 16S and/or 23S rRNA real-time PCR/Pyrosequencing. Compared to culture, PCR/Pyrosequencing displayed 90.9% sensitivity, 99.6% specificity, 95.7% PPV, and 99.1% NPV. Overall concordance rate was 98.9% (1127/1140). In four cases with molecular-positive/culture-negative results, medical chart reviews provided evidence of identical bacteria from subsequent blood or concomitant urine/sputum cultures. Nine culture-positive/molecular-negative cases were associated with either polymicrobial growth, grew only in the anaerobic bottle of the clinical pair, and/or were detected by PCR/Pyrosequencing after 8 hours. In summary, this approach accurately detected and identified bacteria in ~91% of culture-confirmed cases significantly sooner than the phenotypic identification was available, having the potential to improve antibiotic stewardship. PMID:25534615

  16. Aerobic biodegradation of trichloroethene without auxiliary substrates.

    PubMed

    Schmidt, Kathrin R; Gaza, Sarah; Voropaev, Andrey; Ertl, Siegmund; Tiehm, Andreas

    2014-08-01

    Trichloroethene (TCE) represents a priority pollutant and is among the most frequently detected contaminants in groundwater. The current bioremediation measures have certain drawbacks like e.g. the need for auxiliary substrates. Here, the aerobic biodegradation of TCE as the sole growth substrate is demonstrated. This new process of metabolic TCE degradation was first detected in groundwater samples. TCE degradation was stable in an enriched mixed bacterial culture in mineral salts medium for over five years and repeated transfers of the culture resulting in a 10(10) times dilution of the original groundwater. Aerobic TCE degradation resulted in stoichiometric chloride formation. Stable carbon isotope fractionation was observed providing a reliable analytical tool to assess this new biodegradation process at field sites. The results suggest that aerobic biodegradation of TCE without auxiliary substrate could be considered as an option for natural attenuation or engineered bioremediation of contaminated sites. PMID:24793109

  17. CHROMagar Candida Medium for Direct Susceptibility Testing of Yeast from Blood Cultures

    PubMed Central

    Tan, Grace L.; Peterson, Ellena M.

    2005-01-01

    An evaluation was performed on 95 blood cultures positive for Candida spp. to determine the correlation of direct susceptibility testing of fluconazole versus both standardized disk diffusion and MIC methods. For direct testing, an aliquot taken from BD BACTEC Plus and/or BD BACTEC Lytic/10 bottles (Becton Dickinson [BD], Sparks, MD) positive by gram stain for yeast was subcultured to CHROMagar Candida (BD), and a 25-μg fluconazole disk (BD) was placed on the plate. The area of growth inhibition surrounding the disk was measured at 24 and 48 h. In addition, a subculture of the isolate was tested by a microdilution MIC using YeastOne (TREK Diagnostics Systems Inc., OH) and disk diffusion (NCCLS M44-A) using a standardized inoculum plated onto CHROMagar Candida as well as Mueller-Hinton agar to which 2% glucose and 0.5 μg/ml methylene blue dye was added (MH-GMB). The categorical interpretation derived from the MIC was used as the reference to which the disk diffusion results were compared. There were a total of 41 Candida albicans, 23 Candida glabrata, 20 Candida parapsilosis, 9 Candida tropicalis, and 1 each of Candida krusei and Candida lusitaniae tested. At 24 h there was full agreement among the methods for all C. albicans, C. tropicalis, C. lusitaniae, and C. krusei isolates. For the C. parapsilosis isolates at 24 h there was one very major discrepancy using the direct CHROMagar and one major error with the standardized MH-GMB. The majority of the errors were seen at 24 h with the C. glabrata isolates. Of the 23 C. glabrata isolates at 24 h by direct CHROMagar, there were 10 minor and 1 very major error; by MH-GMB there were 12 minor and 2 very major errors; and by standardized CHROMagar Candida there were 13 minor and 2 major errors. There were no very major errors with C. glabrata when all plates were read at 48 h. At 24 h by the direct and standardized CHROMagar the majority of C. glabrata isolates were more resistant, whereas by MH-GMB they were more

  18. Evaluation of a simple blood culture amplification and antigen detection method for diagnosis of Salmonella enterica serovar typhi bacteremia.

    PubMed

    Castonguay-Vanier, Josée; Davong, Viengmon; Bouthasavong, Latsanyphone; Sengdetka, Davanh; Simmalavong, Manivone; Seupsavith, Amphayvanh; Dance, David A B; Baker, Stephen; Le Thi Phuong, Tu; Vongsouvath, Manivanh; Newton, Paul N

    2013-01-01

    In most areas where typhoid is endemic, laboratory diagnosis is not possible due to the lack of appropriate facilities. We investigated whether the combination of blood culture amplification of Salmonella enterica serovar Typhi with an S. Typhi antigen rapid diagnostic test (RDT) could be an accurate and inexpensive tool for the accelerated diagnosis of patients with acute typhoid in Laos. For a panel of 23 Gram-negative reference pathogens, the Standard Diagnostics (catalog no. 15FK20; Kyonggi-do, South Korea) RDT gave positive results for S. Typhi NCTC 8385, S. Typhi NCTC 786 (Vi negative), Salmonella enterica serovar Enteritidis (ATCC 13076), and Salmonella enterica serovar Ndolo NCTC 8700 (all group D). In a prospective study of 6,456 blood culture bottles from 3,028 patients over 15 months, 392 blood culture bottles (6.1%) from 221 (7.3%) patients had Gram-negative rods (GNRs) seen in the blood culture fluid. The sensitivity, negative predictive value, specificity, and positive predictive value were 96.7%, 99.5%, 97.9%, and 87.9%, respectively, for patients with proven S. Typhi bacteremia and 91.2%, 98.4%, 98.9%, and 93.9% for patients with group D Salmonella. The median (range) number of days between diagnosis by RDT and reference assays was 1 (-1 to +2) day for those with confirmed S. Typhi. The use of antigen-based pathogen detection in blood culture fluid may be a useful, relatively rapid, inexpensive, and accurate technique for the identification of important causes of bacteremia in the tropics. PMID:23100346

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

    PubMed

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

    2014-12-01

    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

  20. Cultures and co-cultures of human blood mononuclear cells and endothelial cells for the biocompatibility assessment of surface modified AISI 316L austenitic stainless steel.

    PubMed

    Stio, Maria; Martinesi, Maria; Treves, Cristina; Borgioli, Francesca

    2016-12-01

    Samples of AISI 316L austenitic stainless steel were subjected either to grinding and polishing procedure, or to grinding and then low temperature glow-discharge nitriding treatment, or to grinding, nitriding and subsequently coating with collagen-I. Nitrided samples, even if only ground, show a higher corrosion resistance in PBS solution, in comparison with ground and polished AISI 316L. Biocompatibility was evaluated in vitro by incubating the samples with either peripheral blood mononuclear cells (PBMC) or human umbilical vein endothelial cells (HUVEC), tested separately or in co-culture. HUVEC-PBMC co-culture and co-incubation of HUVEC with PBMC culture medium, after the previous incubation of PBMC with metallic samples, allowed to determine whether the incubation of PBMC with the different samples might affect HUVEC behaviour. Many biological parameters were considered: cell proliferation, release of cytokines, matrix metalloproteinases (MMPs) and sICAM-1, gelatinolytic activity of MMPs, and ICAM-1 protein expression. Nitriding treatment, with or without collagen coating of the samples, is able to ameliorate some of the biological parameters taken into account. The obtained results point out that biocompatibility may be successfully tested in vitro, using cultures of normal human cells, as blood and endothelial cells, but more than one cell line should be used, separately or in co-culture, and different parameters should be determined, in particular those correlated with inflammatory phenomena. PMID:27612806

  1. Management of aerobic vaginitis.

    PubMed

    Tempera, Gianna; Furneri, Pio Maria

    2010-01-01

    Aerobic vaginitis is a new nonclassifiable pathology that is neither specific vaginitis nor bacterial vaginosis. The diversity of this microbiological peculiarity could also explain several therapeutic failures when patients were treated for infections identified as bacterial vaginosis. The diagnosis 'aerobic vaginitis' is essentially based on microscopic examinations using a phase-contrast microscope (at ×400 magnification). The therapeutic choice for 'aerobic vaginitis' should take into consideration an antibiotic characterized by an intrinsic activity against the majority of bacteria of fecal origin, bactericidal effect and poor/absent interference with the vaginal microbiota. Regarding the therapy for aerobic vaginitis when antimicrobial agents are prescribed, not only the antimicrobial spectrum but also the presumed ecological disturbance on the anaerobic and aerobic vaginal and rectal microbiota should be taken into a consideration. Because of their very low impact on the vaginal microbiota, kanamycin or quinolones are to be considered a good choice for therapy. PMID:21051843

  2. Differential sensitivity of aerobic gram-positive and gram-negative microorganisms to 2,4,6-trinitrotoluene (TNT) leads to dissimilar growth and TNT transformation: Results of soil and pure culture studies

    SciTech Connect

    Fuller, M.E.; Manning, J.F. Jr.

    1996-07-30

    The effects of 2,4,6-trinitrotoluene (TNT) on indigenous soil populations and pure bacterial cultures were examined. The number of colony-forming units (CFU) appearing when TNT-contaminated soil was spread on 0.3% molasses plates decreased by 50% when the agar was amended with 67 {mu}g TNT mL{sup -1}, whereas a 99% reduction was observed when uncontaminated soil was plated. Furthermore, TNT-contaminated soil harbored a greater number of organisms able to grow on plates amended with greater than 10 {mu}g TNT mL{sup -1}. The percentage of gram-positive isolates was markedly less in TNT-contaminated soil (7%; 2 of 30) than in uncontaminated soil (61%; 20 of 33). Pseudomonas aeruginosa, Pseudomonas corrugate, Pseudomonasfluorescens and Alcaligenes xylosoxidans made up the majority of the gram-negative isolates from TNT-contaminated soil. Gram-positive isolates from both soils demonstrated marked growth inhibition when greater than 8-16 {mu}g TNT mL{sup -1} was present in the culture media. Most pure cultures of known aerobic gram-negative organisms readily degraded TNT and evidenced net consumption of reduced metabolites. However, pure cultures of aerobic gram-positive bacteria were sensitive to relatively low concentrations of TNT as indicated by the 50% reduction in growth and TNT transformation which was observed at approximately 10 {mu}g TNT mL{sup -1}. Most non-sporeforming gram-positive organisms incubated in molasses media amended with 80 {mu}g TNT mL{sup -1} or greater became unculturable, whereas all strains tested remained culturable when incubated in mineral media amended with 98 {mu}g TNT mL{sup -1}, indicating that TNT sensitivity is likely linked to cell growth. These results indicate that gram-negative organisms are most likely responsible for any TNT transformation in contaminated soil, due to their relative insensitivity to high TNT concentrations and their ability to transform TNT.

  3. Use of Fluorescence In Situ Hybridization for Rapid Identification of Staphylococci in Blood Culture Samples Collected in a Portuguese Hospital ▿ †

    PubMed Central

    Tavares, Ana; Inácio, João; Melo-Cristino, José; Couto, Isabel

    2008-01-01

    Fluorescence in situ hybridization was used for the direct identification of staphylococci in blood cultures collected at a Portuguese hospital where staphylococci account for up to 35% of clinically relevant blood cultures. The assay was able to detect the presence/absence of staphylococci and distinguish Staphylococcus aureus from coagulase-negative staphylococci in 4.5 h. PMID:18562589

  4. Nucleoside transport at the blood-testis barrier studied with primary-cultured sertoli cells.

    PubMed

    Kato, Ryo; Maeda, Tomoji; Akaike, Toshihiro; Tamai, Ikumi

    2005-02-01

    Nucleosides are essential for nucleotide synthesis in testicular spermatogenesis. In the present study, the mechanism of the supply of nucleosides to the testicular system across the blood-testis barrier was studied using primary-cultured Sertoli cells from rats and TM4 cells from mice. Uptake of uridine by these cells was time- and concentration-dependent. Uridine uptake was decreased under Na(+)-free conditions, and the system was presumed to be high affinity, indicating an Na(+)-dependent concentrative nucleoside transporter (CNT) is involved. On the other hand, nitrobenzylthioinosine, a potent inhibitor of Na(+)-independent equilibrative nucleoside transporters (ENTs), inhibited uridine uptake by the Sertoli cells in a concentration-dependent manner. Expression of nucleoside transporters ENT1, ENT2, ENT3, CNT1, CNT2, and CNT3 was detected in Sertoli cells by reverse transcriptase-polymerase chain reaction analysis. Inhibition studies of the uptake of uridine by various nucleosides both in the presence and absence of Na(+) indicated that the most of those expressed nucleoside transporters, ENTs and CNTs, are involved functionally. These results demonstrated that Sertoli cells are equipped with multiple nucleoside transport systems, including ENT1, ENT2, and CNTs, to provide nucleosides for spermatogenesis. PMID:15547112

  5. Selective suppression of cytokine secretion in whole blood cell cultures of patients with colorectal cancer.

    PubMed Central

    Lahm, H.; Schindel, M.; Frikart, L.; Cerottini, J. P.; Yilmaz, A.; Givel, J. C.; Fischer, J. R.

    1998-01-01

    We have investigated the secretion of interferon alpha (IFN-alpha), IFN-gamma, interleukin-1alpha (IL-1alpha), IL-1beta, IL-2 and tumour necrosis factor alpha (TNF-alpha) in whole blood cell cultures (WBCCs) of colorectal cancer patients upon mitogen stimulation. Whereas the values for IL-1beta and TNF-alpha remained virtually unchanged in comparison with healthy control subjects, WBCCs of colorectal cancer patients secreted significantly lower amounts of IFN-alpha (P < 0.005), IFN-gamma (P < 0.0001), IL-1alpha (P < 0.0001) and IL-2 (P < 0.05). This reduction correlated with the progression of the disease. The total leucocyte and monocyte population were almost identical in both groups. In contrast, a dramatic depletion of lymphocytes was observed in colorectal cancer patients, which affected both lymphocyte counts (P < 0.0005) and their distribution (P < 0.0001). Our results suggest a selective suppression of cytokines in colorectal cancer patients that is related to tumour burden. Several mechanisms might account for this phenomenon, one of which might be lymphocyte depletion. PMID:9792144

  6. Antimicrobial resistance trends in blood culture positive Salmonella Paratyphi A isolates from Pondicherry, India.

    PubMed

    Menezes, G A; Harish, B N; Khan, M A; Goessens, W; Hays, J P

    2016-01-01

    Enteric fever is a public health problem with the upsurge in the occurrence of Salmonella isolates that are resistant to ciprofloxacin. In this study, a total of 284 blood culture isolates of S. Paratyphi A were investigated. Of these isolates, 281 (98.9%) were nalidixic acid resistant. A high rate (6.3%) of high-level resistance (≥4 μg/mL) was found to ciprofloxacin. The isolates with ciprofloxacin minimum inhibitory concentrations (MICs) of ≥12 μg/mL had 4 mutations, 2 mutations within the quinolone resistance-determining region of gyrA and 2 mutations also in parC. According to the Clinical Laboratory Standards Institute 2012 MIC breakpoints, 75.0% of isolates were resistant to ciprofloxacin. Finally, 3 major pulsed-field gel electrophoresis patterns were observed among the S. Paratyphi A isolates. The spread of fluoroquinolone resistant S. Paratyphi A necessitates a change toward 'evidence-based' treatment for enteric fever. The research provides a perspective on the increasing prevalence of antimicrobial resistant S. Paratyphi A isolates in this region of India. PMID:27080779

  7. Evaluation of Antimicrobial Therapy of Blood Culture Positive Healthcare-Associated Infections in Children

    PubMed Central

    Vaara, Martti; Anttila, Veli-Jukka; Hoppu, Kalle; Laaksonen, Raisa; Airaksinen, Marja

    2015-01-01

    Aim Knowledge of the quality of antimicrobial therapy (AMT) used for invasive healthcare-associated infections (HAIs) in paediatrics is scarce. Influence of the final information about the isolated pathogen on the subsequent targeted AMT was investigated in our study. Methods Data on 149 children (0–17 years) with blood culture positive HAIs were collected. The causative microbes under investigation were Staphylococcus aureus, Staphylococcus epidermidis, streptococci, Gram negative rods, and mixed infections were likewise included. For adjusting the antimicrobial regimen, an expert panel evaluated the quality of the targeted AMT and the delay of 72 hours after final microbiology results. AMT was regarded as inappropriate if the pathogen was totally resistant to the used antimicrobials (i) or if the chosen therapy was of not optimal efficacy against the pathogen (ii). Results 17% of the patients received inappropriate AMT. Half of these infections 13/26 (50%) were treated with an antimicrobial to which the isolate was resistant. Three (3/13, 23%) of these patients received antimicrobials which were totally ineffective according to in vitro data. Suboptimal or too broad spectrum AMT was administered to 13/26 (50%) patients. The most common causes of inappropriate use were the use of beta-lactams in oxacillin-resistant Staphylococcus epidermidis infections and vancomycin given in oxacillin-sensitive Staphylococcus aureus infections. Conclusion Approximately 17% of the selected cohort received inappropriate AMT. More attention should be paid to the appropriate use of antimicrobials, and training of prescribers should be urgently provided. PMID:26539831

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

    PubMed Central

    Wang, Hye-young; Kim, Sunghyun; Kim, Jungho; Park, Soon-Deok

    2014-01-01

    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 Staphylococcus spp., the nuc gene for S. aureus, and the mecA gene for methicillin resistance. The detection limit of the multiplex real-time PCR assay was 103 CFU/ml per PCR for each gene target. The multiplex real-time PCR assay was evaluated using 118 clinical isolates from various specimen types and a total of 350 positive blood cultures from a continuous monitoring blood culture system. The results obtained with the multiplex real-time PCR assay for the three targets were in agreement with those of conventional identification and susceptibility testing methods except for one organism. Of 350 positive bottle cultures, the sensitivities of the multiplex real-time PCR kit were 100% (166/166 cultures), 97.2% (35/36 cultures), and 99.2% (117/118 cultures) for the 16S rRNA, nuc, and mecA genes, respectively, and the specificities for all three targets were 100%. The Real-MRSA and Real-MRCoNS multiplex real-time PCR assays are very useful for the rapid accurate diagnosis of staphylococcal BSIs. In addition, the Real-MRSA and Real-MRCoNS multiplex real-time PCR assays could have an important impact on the choice of appropriate antimicrobial therapy, based on detection of the mecA gene. PMID:24648566

  9. Anaerobic and aerobic transformation of TNT

    SciTech Connect

    Kulpa, C.F.; Boopathy, R.; Manning, J.

    1996-12-31

    Most studies on the microbial metabolism of nitroaromatic compounds have used pure cultures of aerobic microorganisms. In many cases, attempts to degrade nitroaromatics under aerobic conditions by pure cultures result in no mineralization and only superficial modifications of the structure. However, mixed culture systems properly operated result in the transformation of 2,4,6-trinitrotoluene (TNT) and in some cases mineralization of TNT occurs. In this paper, the mixed culture system is described with emphasis on intermediates and the characteristics of the aerobic microbial process including the necessity for a co-substrate. The possibility of removing TNT under aerobic/anoxic conditions is described in detail. Another option for the biodegradation of TNT and nitroaromatics is under anaerobic, sulfate reducing conditions. In this instance, the nitroaromatic compounds undergo a series of reductions with the formation of amino compounds. TNT under sulfate reducing conditions is reduced to triaminotoluene presumably by the enzyme nitrite reductase, which is commonly found in many Desulfovibrio spp. The removal of nitro groups from TNT is achieved by a series of reductive reactions with the formation of ammonia and toluene by Desulfovibrio sp. (B strain). These metabolic processes could be applied to other nitroaromatic compounds like nitrobenzene, nitrobenzoic acids, nitrophenols, and aniline. The data supporting the anaerobic transformation of TNT under different growth condition are reviewed in this report.

  10. Human Umbilical Cord Blood-Derived Serum for Culturing the Supportive Feeder Cells of Human Pluripotent Stem Cell Lines

    PubMed Central

    Rungsiwiwut, Ruttachuk; Ingrungruanglert, Praewphan; Numchaisrika, Pranee; Virutamasen, Pramuan; Phermthai, Tatsanee; Pruksananonda, Kamthorn

    2016-01-01

    Although human pluripotent stem cells (hPSCs) can proliferate robustly on the feeder-free culture system, genetic instability of hPSCs has been reported in such environment. Alternatively, feeder cells enable hPSCs to maintain their pluripotency. The feeder cells are usually grown in a culture medium containing fetal bovine serum (FBS) prior to coculture with hPSCs. The use of FBS might limit the clinical application of hPSCs. Recently, human cord blood-derived serum (hUCS) showed a positive effect on culture of mesenchymal stem cells. It is interesting to test whether hUCS can be used for culture of feeder cells of hPSCs. This study was aimed to replace FBS with hUCS for culturing the human foreskin fibroblasts (HFFs) prior to feeder cell preparation. The results showed that HFFs cultured in hUCS-containing medium (HFF-hUCS) displayed fibroblastic features, high proliferation rates, short population doubling times, and normal karyotypes after prolonged culture. Inactivated HFF-hUCS expressed important genes, including Activin A, FGF2, and TGFβ1, which have been implicated in the maintenance of hPSC pluripotency. Moreover, hPSC lines maintained pluripotency, differentiation capacities, and karyotypic stability after being cocultured for extended period with inactivated HFF-hUCS. Therefore, the results demonstrated the benefit of hUCS for hPSCs culture system. PMID:26839561

  11. Blood Culture and Stimulation Conditions for the Diagnosis of Tuberculosis in Cervids by the Cervigam Assay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mitogen and antigen induced interferon-gamma (IFN-gamma) responses of peripheral blood leukocytes from cervids were evaluated using a commercial, whole blood assay for the cytokine (Cervigam trademark, Prionics AG). Whole blood was from Mycobacterium bovis-infected white-tailed deer and reindeer, M....

  12. Methods to determine aerobic endurance.

    PubMed

    Bosquet, Laurent; Léger, Luc; Legros, Patrick

    2002-01-01

    data instead of blood lactate concentration ([La(-)](b)) is not valid. Added to the fact that the AT may not represent the optimal training intensity for elite athletes, it raises doubt on the usefulness of this theory without questioning, however, the usefulness of the whole [La(-)](b)-power curve to assess aerobic endurance and predict performance in long-distance events. PMID:12196030

  13. Teaching Aerobic Lifestyles: New Perspectives.

    ERIC Educational Resources Information Center

    Goodrick, G. Ken; Iammarino, Nicholas K.

    1982-01-01

    New approaches to teaching aerobic life-styles in secondary schools are suggested, focusing on three components: (1) the psychological benefits of aerobic activity; (2) alternative aerobic programs at nonschool locations; and (3) the development of an aerobics curriculum to help maintain an active life-style after graduation. (JN)

  14. Cost-Effectiveness of 30- Compared to 20-Milliliter Blood Cultures: a Retrospective Study

    PubMed Central

    Cheruvanky, Anita; Kirn, Thomas J.

    2015-01-01

    The importance of blood culture (BC) volume for detection of bloodstream infections (BSIs) is documented. Recently, improved diagnostic sensitivity was demonstrated for 30- versus 20-ml BCs in adults (Cockerill FR, Wilson JW, Vetter EA, Goodman KM, Torgerson CA, Harmsen WS, Schleck CD, IIstrup DM, Washington JA, Wilson WR. Clin Infect Dis 38:1724–1730, 2004, http://dx.doi.org/10.1128/JCM.01314-11). Hospitals receive higher reimbursement for patients with documented septicemia. We determined the cost-effectiveness of 30-ml versus 20-ml BCs using results from our institution and previously published data. Positive BC results from 292 bacteremic episodes were reviewed. The costs of the reagents, equipment, phlebotomist, and technologist time were determined. The medical records department provided Medicare reimbursement (MR) data for patients with selected ICD-9 codes. These data provided an estimate of the annualized increase in MR versus costs associated with conversion to 30-ml BCs. MR for 464 annual primary BSIs was $24,808/episode. An expected 7.2% increase in BSIs detected using 30-ml BCs would add 34 additional cases annually and increase MR by $843,472. Comparative MR data for cases where septicemia complicated another diagnosis were available for 4 International Classification of Diseases, Ninth Revision (ICD-9) codes: laparoscopic cholecystectomy, biliary tract disorders, pneumonia, and cellulitis. The mean incremental MR was $9,667 per episode, which projected to a $483,350 revenue increase annually. The annual cost associated with conversion to 30-ml BCs was estimated to be $157,798. Thus, the potential net increase in hospital revenue would be $1,169,031 for 30-ml versus 20-ml BCs. Our results suggest that conversion to 30-ml BCs may not only improve patient care by detecting more BSIs but also increase hospital revenue substantially. PMID:26491177

  15. Cost-Effectiveness of 30- Compared to 20-Milliliter Blood Cultures: a Retrospective Study.

    PubMed

    Cheruvanky, Anita; Kirn, Thomas J; Weinstein, Melvin P

    2016-01-01

    The importance of blood culture (BC) volume for detection of bloodstream infections (BSIs) is documented. Recently, improved diagnostic sensitivity was demonstrated for 30- versus 20-ml BCs in adults (Cockerill FR, Wilson JW, Vetter EA, Goodman KM, Torgerson CA, Harmsen WS, Schleck CD, IIstrup DM, Washington JA, Wilson WR. Clin Infect Dis 38:1724-1730, 2004, http://dx.doi.org/10.1128/JCM.01314-11). Hospitals receive higher reimbursement for patients with documented septicemia. We determined the cost-effectiveness of 30-ml versus 20-ml BCs using results from our institution and previously published data. Positive BC results from 292 bacteremic episodes were reviewed. The costs of the reagents, equipment, phlebotomist, and technologist time were determined. The medical records department provided Medicare reimbursement (MR) data for patients with selected ICD-9 codes. These data provided an estimate of the annualized increase in MR versus costs associated with conversion to 30-ml BCs. MR for 464 annual primary BSIs was $24,808/episode. An expected 7.2% increase in BSIs detected using 30-ml BCs would add 34 additional cases annually and increase MR by $843,472. Comparative MR data for cases where septicemia complicated another diagnosis were available for 4 International Classification of Diseases, Ninth Revision (ICD-9) codes: laparoscopic cholecystectomy, biliary tract disorders, pneumonia, and cellulitis. The mean incremental MR was $9,667 per episode, which projected to a $483,350 revenue increase annually. The annual cost associated with conversion to 30-ml BCs was estimated to be $157,798. Thus, the potential net increase in hospital revenue would be $1,169,031 for 30-ml versus 20-ml BCs. Our results suggest that conversion to 30-ml BCs may not only improve patient care by detecting more BSIs but also increase hospital revenue substantially. PMID:26491177

  16. Utilization of blood cultures in Danish hospitals: a population-based descriptive analysis.

    PubMed

    Gubbels, S; Nielsen, J; Voldstedlund, M; Kristensen, B; Schønheyder, H C; Vandenbroucke-Grauls, C M J E; Arpi, M; Björnsdóttir, M K; Knudsen, J Dahl; Dessau, R B; Jensen, T Gorm; Kjældgaard, P; Lemming, L; Møller, J K; Hansen, D Schrøder; Mølbak, K

    2015-04-01

    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 of nationwide surveillance for bacteraemia. From the Danish Microbiology Database, we retrieved all BCs taken between 2010 and 2013 and linked these to admission data from the National Patient Registry. In total, 4 587 295 admissions were registered, and in 11%, at least one BC was taken. Almost 50% of BCs were taken at admission. The chance of having a BC taken declined over the next days but increased after 4 days of admission. Data linkage identified 876 290 days on which at least one BC was taken; 6.4% yielded positive results. Ten species, Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, Streptococcus pneumoniae, Enterococcus faecium, Enterococcus faecalis, Pseudomonas aeruginosa, Candida albicans, Enterobacter cloacae and Klebsiella oxytoca, accounted for 74.7% of agents for this purpose classified as pathogenic. An increase in BCs and positive BCs was observed over time, particularly among older patients. BCs showed a seasonal pattern overall and for S. pneumoniae particularly. A predominance of male patients was seen for bacteraemias due to S. aureus, E. faecium and K. pneumoniae. Minor differences in BCs and positive BCs between departments of clinical microbiology underpin the rationale of a future automated surveillance for bacteraemia. The study also provides important knowledge for interpretation of surveillance of invasive infections more generally. PMID:25658520

  17. Aerobic Conditioning Class.

    ERIC Educational Resources Information Center

    Johnson, Neil R.

    1980-01-01

    An aerobic exercise class that focuses on the conditioning of the cardiovascular and muscular systems is presented. Students complete data cards on heart rate, pulse, and exercises to be completed during the forty minute course. (CJ)

  18. Data correction pre-processing for electronically stored blood culture results: Implications on microbial spectrum and empiric antibiotic therapy

    PubMed Central

    2009-01-01

    Background The outcome of patients with bacteraemia is influenced by the initial selection of adequate antimicrobial therapy. The objective of our study was to clarify the influence of different crude data correction methods on a) microbial spectrum and ranking of pathogens, and b) cumulative antimicrobial susceptibility pattern of blood culture isolates obtained from patients from intensive care units (ICUs) using a computer based tool, MONI. Methods Analysis of 13 ICUs over a period of 7 years yielded 1427 microorganisms from positive results. Three different data correction methods were applied. Raw data method (RDM): Data without further correction, including all positive blood culture results. Duplicate-free method (DFM): Correction of raw data for consecutive patient's results yielding same microorganism with similar antibiogram within a two-week period. Contaminant-free method (CFM): Bacteraemia caused by possible contaminants was only assumed as true bloodstream infection, if an organism of the same species was isolated from > 2 sets of blood cultures within 5 days. Results Our study demonstrates that different approaches towards raw data correction – none (RDM), duplicate-free (DFM), and a contaminant-free method (CFM) – show different results in analysis of positive blood cultures. Regarding the spectrum of microorganisms, RDM and DFM yielded almost similar results in ranking of microorganisms, whereas using the CFM resulted in a clinically and epidemiologically more plausible spectrum. Conclusion For possible skin contaminants, the proportion of microorganisms in terms of number of episodes is most influenced by the CFM, followed by the DFM. However, with exception of fusidic acid for gram-positive organisms, none of the evaluated correction methods would have changed advice for empiric therapy on the selected ICUs. PMID:19500418

  19. Draft Genome Sequence of “Terrisporobacter othiniensis” Isolated from a Blood Culture from a Human Patient

    PubMed Central

    Lund, Lars Christian; Sydenham, Thomas Vognbjerg; Høgh, Silje Vermedal; Skov, Marianne; Kemp, Michael

    2015-01-01

    “Terrisporobacter othiniensis” (proposed species) was isolated from a blood culture. Genomic DNA was sequenced using a MiSeq benchtop sequencer (Illumina) and assembled using the SPAdes genome assembler. This resulted in a draft genome sequence comprising 3,980,019 bp in 167 contigs containing 3,449 coding sequences, 7 rRNAs, and 58 tRNAs. PMID:25744994

  20. Detection of Legionella pneumophila serogroup 1 in blood cultures from a patient treated with tumor necrosis factor-alpha inhibitor.

    PubMed

    Kaku, Norihito; Yanagihara, Katsunori; Morinaga, Yoshitomo; Sato, Tsuyoshi; Nakashima, Munetoshi; Sakai, Takahiro; Tominaga, Hiroo; Wakigawa, Fumiko; Nagashima, Seiji; Fukuda, Minoru; Hashiguchi, Kohji; Kohno, Shigeru

    2013-02-01

    A 65-year-old man was admitted to our hospital with a temperature of 39.3 °C, cough, sputum, and pharyngeal discomfort that had persisted for 3 days. He had been treated with methotrexate and adalimumab (a tumor necrosis factor-alpha [TNF-α] inhibitor) for rheumatoid arthritis for 2 years, and he had also been treated with S-1 (tegafur, gimeracil, and oteracil potassium) for pancreatic metastasis of gastric cancer for 2 months. Regardless of the underlying pathologies, his general condition was good and he had worked as an electrician until 2 days before admission. However, his appetite had suddenly decreased from the day before admission, and high fever and hypoxia were also evident upon admission. A chest X-ray and computed tomography scan revealed left pleural effusion and consolidation in both lungs. The pneumonia severity index score was 165 and the risk class was V. Accordingly, we started to treat the pneumonia with a combination of levofloxacin and meropenem. Thereafter, we received positive urinary antigen test findings for Legionella pneumophila. After hospitalization, hypoxia was progressed and hypotension was emerged. Despite the application of appropriate antibiotics, vasopressors, and oxygenation, the patient died 8 h after admission. Even after his death, blood cultures were continued to consider the possibility of bacterial co-infection. Although no bacteria were detected from blood cultures, Gimenez staining revealed pink bacteria in blood culture fluids. Subsequent blood fluid culture in selective medium revealed L. pneumophila serogroup 1. Recently, TNF-α inhibitors have been described as a risk factor for Legionnaires' disease. In consideration of the increased frequency of TNF-α inhibitors, we may need to recognize anew that L. pneumophila might be a pathogen of severe community-acquired pneumonia. PMID:22911089

  1. Time-to-reporting of blood culture positivity and central venous catheter-associated Candida glabrata fungemia in cancer patients.

    PubMed

    Stempel, Jessica M; Farmakiotis, Dimitrios; Tarrand, Jeffrey J; Kontoyiannis, Dimitrios P

    2016-07-01

    Among cancer patients with Candida glabrata (the Candida species with the slowest in-vitro growth) fungemia, time-to-positive blood culture reporting (TTR) was shorter in catheter-associated candidemia (mean±standard deviation: 67±35 h) than in candidemia from other sources (79±31, P<.01). TTR<48 h was 92% specific for catheter-associated C. glabrata fungemia. PMID:27133559

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

    PubMed

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

    2015-02-01

    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

  3. Application of the Accurate Mass and Time Tag Approach to the Proteome Analysis of Sub-cellular Fractions Obtained from Rhodobacter sphaeroides 2.4.1 Aerobic and Photosynthetic Cell Cultures

    SciTech Connect

    Callister, Stephen J.; Dominguez, Migual; Nicora, Carrie D.; Zeng, Xiaohua; Tavano, Christine; Kaplan, Samuel; Donohue, Timothy; Smith, Richard D.; Lipton, Mary S.

    2006-08-04

    Abstract The high-throughput accurate mass and time tag (AMT) proteomic approach was utilized to characterize the proteomes for cytoplasm, cytoplasmic membrane, periplasm, and outer membrane fractions from aerobic and photosynthetic cultures of the gram-nagtive bacterium Rhodobacter sphaeroides 2.4.1. In addition, we analyzed the proteins within purified chromatophore fractions that house the photosynthetic apparatus from photosynthetically grown cells. In total, 8300 peptides were identified with high confidence from at least one sub-cellular fraction from either cell culture. These peptides were derived from 1514 genes or 35% percent of proteins predicted to be encoded by the genome. A significant number of these proteins were detected within a single sub-cellular fraction and their localization was compared to in-silico predictions. However, the majority of proteins were observed in multiple sub-cellular fractions, and the most likely sub-cellular localization for these proteins was investigated using a Z-score analysis of peptide abundance along with clustering techniques. Good (81%) agreement was observed between the experimental results and in-silico predictions. The AMT tag approach provides localization evidence for those proteins that have no predicted localization information, those annotated as putative proteins, and/or for those proteins annotated as hypothetical and conserved hypothetical.

  4. Summary report on the aerobic degradation of diesel fuel and the degradation of toluene under aerobic, denitrifying and sulfate reducing conditions

    SciTech Connect

    Coyne, P.; Smith, G.

    1995-08-15

    This report contains a number of studies that were performed to better understand the technology of the biodegradation of petroleum hydrocarbons. Topics of investigation include the following: diesel fuel degradation by Rhodococcus erythropolis; BTEX degradation by soil isolates; aerobic degradation of diesel fuel-respirometry; aerobic degradation of diesel fuel-shake culture; aerobic toluene degradation by A3; effect of HEPES, B1, and myo-inositol addition on the growth of A3; aerobic and anaerobic toluene degradation by contaminated soils; denitrifying bacteria MPNs; sulfate-reducing bacteria MPNs; and aerobic, DNB and SRB enrichments.

  5. Bacteriological Profile and Drug Resistance Patterns of Blood Culture Isolates in a Tertiary Care Nephrourology Teaching Institute

    PubMed Central

    Gang, Sishir; Sabnis, Ravindra; Desai, Mahesh

    2014-01-01

    Blood stream infections can lead to life threatening sepsis and require rapid antimicrobial treatment. The organisms implicated in these infections vary with the geographical alteration. Infections caused by MDR organisms are more likely to increase the risk of death in these patients. The present study was aimed to study the profile of organisms causing bacteremia and understand antibiotic resistance patterns in our hospital. 1440 blood samples collected over a year from clinically suspected cases of bacteremia were studied. The isolates were identified by standard biochemical tests and antimicrobial resistance patterns were determined by CLSI guidelines. Positive blood cultures were obtained in 9.2% of cases of which Gram-positive bacteria accounted for 58.3% of cases with staph aureus predominance; gram negative bacteria accounted for 40.2% with enterobactereciea predominence; and 1.5% were fungal isolates. The most sensitive drugs for Gram-positive isolates were vancomycin, teicoplanin, daptomycin, linezolid, and tigecycline and for Gram-negative were carbapenems, colistin, aminoglycosides, and tigecycline. The prevalence of MRSA and vancomycin resistance was 70.6% and 21.6%, respectively. ESBL prevalence was 39.6%. Overall low positive rates of blood culture were observed. PMID:24804199

  6. Controlled Comparative Evaluation of BacT/Alert FAN and ESP 80A Aerobic Media as Means for Detecting Bacteremia and Fungemia

    PubMed Central

    Doern, Gary V.; Barton, Ann; Rao, Sudah

    1998-01-01

    During a one-year period, a total of 6,305 blood cultures were processed in a tertiary-care teaching hospital; 6 to 12 ml of blood was inoculated into both a BacT/Alert Fan aerobic bottle and an ESP 80A aerobic bottle. The FAN aerobic bottle contains an antimicrobial-absorbing material; the 80A aerobic bottle does not. Bottles were processed on their respective continuous-monitoring blood culture instruments for up to five days of incubation. Four hundred thirty-three cultures (6.9%) representing 301 septic episodes in 235 different patients yielded 490 bacteria or yeasts thought to be clinically significant. Two hundred seventy-five of the 433 presumed clinically significant positive cultures (63.5%) representing 195 septic episodes and yielding 301 isolates were positive in both FAN and 80A bottles. One hundred nine significant positive cultures (25.2%) (i.e., cultures positive with an organism judged to be of probable clinical significance) from 70 septic episodes yielded 126 isolates only in FAN bottles. Conversely, the 80A bottle was exclusively positive in 49 instances (11.3%), representing 36 septic episodes and yielding 63 isolates. The higher rates of significant positive blood cultures, numbers of septic episodes documented, and numbers of isolates recovered in FAN bottles versus 80A bottles were all statistically significant (P < 0.05). Enhanced rates of detection of presumed clinically significant isolates in FAN bottles were largely accounted for by Staphylococcus aureus, members of the Enterobacteriaceae, and non-Pseudomonas aeruginosa miscellaneous gram-negative bacilli from patients receiving antimicrobial therapy at the time blood cultures were obtained. Enhanced recovery of one organism group, the β-hemolytic streptococci, occurred in 80A. With one exception, detection times were essentially equivalent in the two systems. The single exception pertained to streptococci and enterococci, which were recovered significantly faster in 80A bottles

  7. Development of a Xeno-Free Autologous Culture System for Endothelial Progenitor Cells Derived from Human Umbilical Cord Blood

    PubMed Central

    Park, Soon-Jung; Kim, Hojin; Bae, Daekyeong

    2013-01-01

    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. PMID:24086472

  8. PRIMARY CULTURE OF CHOROIDAL EPITHELIAL CELLS: CHARACTERIZATION OF AN IN VITRO MODEL OF BLOOD-CSF BARRIER

    PubMed Central

    ZHENG, WEI; ZHAO, QIUQU; GRAZIANO, JOSEPH H.

    2016-01-01

    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

  9. Evaluation of the BD Max StaphSR Assay for Rapid Identification of Staphylococcus aureus and Methicillin-Resistant S. aureus in Positive Blood Culture Broths

    PubMed Central

    Hofko, Marjeta; Hamilton, Fiona; Mackenzie, Laura; Zimmermann, Stefan; Templeton, Kate

    2015-01-01

    We evaluated the performance of the BD Max StaphSR assay for the direct detection of Staphylococcus aureus from blood culture medium. In a two-center trial, 155 blood cultures from the BD Bactec FX system and 212 from the bioMérieux BacT/Alert system were tested; 170 bottles yielded S. aureus, and all were identified correctly by the BD Max StaphSR assay. The assay required approximately 2.5 h, thus allowing rapid identification of blood cultures flagged positive. PMID:26292311

  10. Comparison of the TEMPO® System, Petrifilm® , and Cultural MPN Procedure for Enumeration of E. coli, Coliforms and Total Aerobic Plate Counts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction: Recent innovations in microbiological methods for analysis of food products have been in methods for detection of bacterial pathogens. Petrifilm dehydrated plates are the only significant addition to cultural procedures for indicator organisms in the last 20 years. An automated most...

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

    PubMed Central

    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.

    2016-01-01

    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

  12. Development of a selective culture medium for bifidobacteria, Raffinose-Propionate Lithium Mupirocin (RP-MUP) and assessment of its usage with Petrifilm™ Aerobic Count plates.

    PubMed

    Miranda, Rodrigo Otávio; de Carvalho, Antonio Fernandes; Nero, Luís Augusto

    2014-05-01

    This study aimed to develop a selective culture media to enumerate bifidobacteria in fermented milk and to assess this medium when used with Petrifilm™ AC plates. For this purpose, Bifidobacterium spp., Lactobacillus spp. and Streptococcus thermophilus strains were tested to verify their fermentation patterns for different carbohydrates. All bifidobacteria strains were able to use raffinose. Based on these characteristic, a selective culture medium was proposed (Raffinose-Propionate Lithium Mupirocin, RP-MUP), used with Petrifilm™ AC plates, and was used to enumerate bifidobacteria in fermented milk. RP-MUP performance was assessed by comparing the results with this medium to reference protocols and culture media for bifidobacteria enumeration. RP-MUP, whether used or not with Petrifilm™ AC, presented similar performance to TOS-MUP (ISO 29981), with no significant differences between the mean bifidobacteria counts (p < 0.05) and with high correlation indices (r = 0.99, p < 0.05). As an advantage, reliable results were obtained after just 48 h of incubation when RP-MUP was used with Petrifilm™ AC, instead of the 72 h described in the ISO 29981 protocol. PMID:24387858

  13. Sexual dimorphism in primate aerobic capacity: a phylogenetic test.

    PubMed

    Lindenfors, Patrik; Revell, L J; Nunn, C L

    2010-06-01

    Male intrasexual competition should favour increased male physical prowess. This should in turn result in greater aerobic capacity in males than in females (i.e. sexual dimorphism) and a correlation between sexual dimorphism in aerobic capacity and the strength of sexual selection among species. However, physiological scaling laws predict that aerobic capacity should be lower per unit body mass in larger than in smaller animals, potentially reducing or reversing the sex difference and its association with measures of sexual selection. We used measures of haematocrit and red blood cell (RBC) counts from 45 species of primates to test four predictions related to sexual selection and body mass: (i) on average, males should have higher aerobic capacity than females, (ii) aerobic capacity should be higher in adult than juvenile males, (iii) aerobic capacity should increase with increasing sexual selection, but also that (iv) measures of aerobic capacity should co-vary negatively with body mass. For the first two predictions, we used a phylogenetic paired t-test developed for this study. We found support for predictions (i) and (ii). For prediction (iii), however, we found a negative correlation between the degree of sexual selection and aerobic capacity, which was opposite to our prediction. Prediction (iv) was generally supported. We also investigated whether substrate use, basal metabolic rate and agility influenced physiological measures of oxygen transport, but we found only weak evidence for a correlation between RBC count and agility. PMID:20406346

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

    SciTech Connect

    Weinstein, M.P.; Mirrett, S.; Reller, L.B.

    1988-05-01

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

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

    PubMed

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

    2016-07-01

    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 (p<0.001) decrease in the percentage of CD3(+) T lymphocytes within the PBMCs was found. More specifically, this reduction was due to a significant (p<0.01) decrease in the percentage of γδ(+) T lymphocytes. Thus measuring immune responses on purified PBMCs might not give a physiologically relevant output. Additionally, it was tested if the 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

  16. Functionalized arrays of Raman-enhancing nanoparticles for capture and culture-free analysis of bacteria in human blood

    NASA Astrophysics Data System (ADS)

    Liu, Ting-Yu; Tsai, Kun-Tong; Wang, Huai-Hsien; Chen, Yu; Chen, Yu-Hsuan; Chao, Yuan-Chun; Chang, Hsuan-Hao; Lin, Chi-Hung; Wang, Juen-Kai; Wang, Yuh-Lin

    2011-11-01

    Detecting bacteria in clinical samples without using time-consuming culture processes would allow rapid diagnoses. Such a culture-free detection method requires the capture and analysis of bacteria from a body fluid, which are usually of complicated composition. Here we show that coating Ag-nanoparticle arrays with vancomycin (Van) can provide label-free analysis of bacteria via surface-enhanced Raman spectroscopy (SERS), leading to a ~1,000-fold increase in bacteria capture, without introducing significant spectral interference. Bacteria from human blood can be concentrated onto a microscopic Van-coated area while blood cells are excluded. Furthermore, a Van-coated substrate provides distinctly different SERS spectra of Van-susceptible and Van-resistant Enterococcus, indicating its potential use for drug-resistance tests. Our results represent a critical step towards the creation of SERS-based multifunctional biochips for rapid culture- and label-free detection and drug-resistant testing of microorganisms in clinical samples.

  17. A Triple Culture Model of the Blood-Brain Barrier Using Porcine Brain Endothelial cells, Astrocytes and Pericytes

    PubMed Central

    Thomsen, Louiza Bohn; Burkhart, Annette; Moos, Torben

    2015-01-01

    In vitro blood-brain barrier (BBB) models based on primary brain endothelial cells (BECs) cultured as monoculture or in co-culture with primary astrocytes and pericytes are useful for studying many properties of the BBB. The BECs retain their expression of tight junction proteins and efflux transporters leading to high trans-endothelial electric resistance (TEER) and low passive paracellular permeability. The BECs, astrocytes and pericytes are often isolated from small rodents. Larger species as cows and pigs however, reveal a higher yield, are readily available and have a closer resemblance to humans, which make them favorable high-throughput sources for cellular isolation. The aim of the present study has been to determine if the preferable combination of purely porcine cells isolated from the 6 months old domestic pigs, i.e. porcine brain endothelial cells (PBECs) in co-culture with porcine astrocytes and pericytes, would compare with PBECs co-cultured with astrocytes and pericytes isolated from newborn rats with respect to TEER value and low passive permeability. The astrocytes and pericytes were grown both as contact and non-contact co-cultures as well as in triple culture to examine their effects on the PBECs for barrier formation as revealed by TEER, passive permeability, and expression patterns of tight junction proteins, efflux transporters and the transferrin receptor. This syngenic porcine in vitro BBB model is comparable to triple cultures using PBECs, rat astrocytes and rat pericytes with respect to TEER formation, low passive permeability, and expression of hallmark proteins signifying the brain endothelium (tight junction proteins claudin 5 and occludin, the efflux transporters P-glycoprotein (PgP) and breast cancer related protein (BCRP), and the transferrin receptor). PMID:26241648

  18. Impact of a Culturally Sensitive Health Self-Empowerment Workshop Series on Health Behaviors/Lifestyles, BMI, and Blood Pressure of Culturally Diverse Overweight/Obese Adults

    PubMed Central

    Tucker, Carolyn M.; Butler, Ashley; Kaye, Lillian B.; Nolan, Sarah E. M.; Flenar, Delphia J.; Marsiske, Michael; Bragg, Marie; Hoover, Eddie; Daly, Katherine

    2013-01-01

    Objective Examine the impact of the Health Self-Empowerment Theory-based, culturally sensitive Health Self-Empowerment (HSE) Workshop Series to Modify and Prevent Obesity on levels of health promoting (health-smart) behaviors, motivators of and barriers to these behaviors, health promoting lifestyle variables, and health status indicators (Body Mass Index [BMI] and blood pressure) among a culturally diverse sample of overweight/obese adults from mostly low income households. Design 153 overweight/obese adults participated in an Immediate Treatment (IT) Group (n = 100) or a Waitlist Control (WC) Group (n = 53). Results Post-intervention, the IT Group compared to the WC Group reported (a) significantly higher engagement in physical activity and healthy eating, (b) significantly less intake of calories, total fat, transfat, saturated fat, sugar, and added sugar, (c) significantly higher motivators for engaging in two of four specific health-smart behaviors, (d) significantly lower barriers to engaging in three of four specific health-smart behaviors, and (e) significantly lower BMI and systolic blood pressure. Conclusion The HSE Workshop Series may be an effective intervention for treating and preventing obesity among diverse low-income adults – individuals who often perceive/experience limited power over their health. Health care providers, particularly physicians, have important health empowerment roles in this intervention. PMID:24910589

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

    PubMed

    Celik, Ayla

    2006-01-01

    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

  20. Dance--Aerobic and Anaerobic.

    ERIC Educational Resources Information Center

    Cohen, Arlette

    1984-01-01

    This article defines and explains aerobic exercise and its effects on the cardiovascular system. Various studies on dancers are cited indicating that dance is an anaerobic activity with some small degree of aerobic benefit. (DF)

  1. Microarray technology for yeast identification directly from positive blood cultures. A multicenter Italian experience.

    PubMed

    Farina, Claudio; Russello, Giuseppe; Andreoni, Stefano; Bonetti, Cristina; Conte, Marco; Fazi, Paolo; Lombardi, Gianluigi; Luzzaro, Francesco; Manso, Esther; Marone, Piero; Passera, Marco; Rocchetti, Andrea; Sanna, Silvana; Viganò, Egidio Franco

    2012-07-01

    The authors evaluated the performance of the MycArray™ Yeast ID (Myconostica Ltd, UK) assay in the identification of a total of 88 yeast isolates recovered in culture as compared to that obtained through routine methods. The turn-around time for species identification directly from cultures by the MycArray was 6 hours, much quicker than classical methods and all yeasts were correctly identified. In two cases a double identification including Saccharomyces cerevisiae was noted, but it was not confirmed by culture. The results show that MycArray Yeast ID can be a potential tool for rapid detection and identification of Candida species. PMID:22217211

  2. Agreement of Direct Antifungal Susceptibility Testing from Positive Blood Culture Bottles with the Conventional Method for Candida Species.

    PubMed

    Jabeen, Kauser; Kumar, Haresh; Farooqi, Joveria; Mehboob, Raunaq; Brandt, Mary E; Zafar, Afia

    2016-02-01

    Early availability of antifungal susceptibilities can ensure timely institution of targeted therapy in candidemia, which can improve patient outcomes. This study prospectively determines the agreement between the results of direct testing of antifungal susceptibilities from blood culture bottles by disk diffusion and Etest and the results of standardized susceptibility testing methods; direct testing would allow susceptibility results to be available 1 to 2 days earlier. A total of 104 blood cultures with different Candida species (28% C. albicans, 27% C. parapsilosis, 26% C. tropicalis, etc.) were evaluated between January 2012 and May 2013 for agreement of fluconazole, voriconazole, and amphotericin B susceptibility results by disk diffusion. Agreement in MICs obtained by Etest was determined for fluconazole (21 isolates), voriconazole (28 isolates), amphotericin (29 isolates), and caspofungin (29 isolates). The kappa scores for categorical agreement were highest for fluconazole by disk diffusion (0.902, standard error [SE] = 0.076) and Etest (1.00, SE = 0.218) and for amphotericin B by disk diffusion (1.00, SE = 0.098). The Pearson correlation (r) of zone diameters was strongest for fluconazole (0.69) and amphotericin (0.70) and moderate for voriconazole (0.60), and the Pearson correlation of MICs was strongest for fluconazole (0.94) and caspofungin (0.88). However, the moderate correlation of amphotericin MICs with zone diameters (-0.42) precludes the use of amphotericin B disk diffusion for susceptibility testing. There were no very major errors; however, there were 1 (1%) major and 5 (4.8%) minor errors with disk diffusion and 4 (13.3%) minor errors with Etest. Thus, antifungal disk diffusion directly from blood culture bottles is a rapid and easy method for fluconazole and voriconazole susceptibility testing for timely tailoring of candidemia therapy. PMID:26607985

  3. Activities of potential therapeutic and prophylactic antibiotics against blood culture isolates of viridans group streptococci from neutropenic patients receiving ciprofloxacin.

    PubMed Central

    McWhinney, P H; Patel, S; Whiley, R A; Hardie, J M; Gillespie, S H; Kibbler, C C

    1993-01-01

    All 47 sequential blood culture isolates of viridans group streptococci obtained from febrile neutropenic patients receiving quinolone prophylaxis were susceptible to vancomycin, teicoplanin, and imipenem. Resistance to benzylpenicillin (MIC for 50% of isolates [MIC50], 0.125 microgram/ml) and ceftazidime (MIC50, 4 micrograms/ml) was common. Most isolates were susceptible to amoxicillin, co-amoxiclav (amoxicillin-clavulanic acid at a 2:1 ratio by weight), azlocillin, clarithromycin, and erythromycin, with azithromycin showing comparable activity. The MIC90 of sparfloxacin was 1 microgram/ml; those for ciprofloxacin and ofloxacin were > 16 and 16 micrograms/ml, respectively. PMID:8285642

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

    PubMed

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

    2016-03-01

    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

  5. Evaluation of a Fully Automated Research Prototype for the Immediate Identification of Microorganisms from Positive Blood Cultures under Clinical Conditions

    PubMed Central

    Hyman, Jay M.; Walsh, John D.; Ronsick, Christopher; Wilson, Mark; Hazen, Kevin C.; Borzhemskaya, Larisa; Link, John; Clay, Bradford; Ullery, Michael; Sanchez-Illan, Mirta; Rothenberg, Steven; Robinson, Ron; van Belkum, Alex

    2016-01-01

    ABSTRACT A clinical laboratory evaluation of an intrinsic fluorescence spectroscopy (IFS)-based identification system paired to a BacT/Alert Virtuo microbial detection system (bioMérieux, Inc., Durham, NC) was performed to assess the potential for fully automated identification of positive blood cultures. The prototype IFS system incorporates a novel method combining a simple microbial purification procedure with rapid in situ identification via spectroscopy. Results were available within 15 min of a bottle signaling positive and required no manual intervention. Among cultures positive for organisms contained within the database and producing acceptable spectra, 75 of 88 (85.2%) and 79 of 88 (89.8%) were correctly identified to the species and genus level, respectively. These results are similar to the performance of existing rapid methods. PMID:27094332

  6. Real-Time Identification of Bacteria and Candida Species in Positive Blood Culture Broths by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry▿

    PubMed Central

    Ferroni, Agnès; Suarez, Stéphanie; Beretti, Jean-Luc; Dauphin, Brunhilde; Bille, Emmanuelle; Meyer, Julie; Bougnoux, Marie-Elisabeth; Alanio, Alexandre; Berche, Patrick; Nassif, Xavier

    2010-01-01

    Delays in the identification of microorganisms are a barrier to the establishment of adequate empirical antibiotic therapy of bacteremia. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) allows the identification of microorganisms directly from colonies within minutes. In this study, we have adapted and tested this technology for use with blood culture broths, thus allowing identification in less than 30 min once the blood culture is detected as positive. Our method is based on the selective recovery of bacteria by adding a detergent that solubilizes blood cells but not microbial membranes. Microorganisms are then extracted by centrifugation and analyzed by MALDI-TOF-MS. This strategy was first tested by inoculating various bacterial and fungal species into negative blood culture bottles. We then tested positive patient blood or fluid samples grown in blood culture bottles, and the results obtained by MALDI-TOF-MS were compared with those obtained using conventional strategies. Three hundred twelve spiked bottles and 434 positive cultures from patients were analyzed. Among monomicrobial fluids, MALDI-TOF-MS allowed a reliable identification at the species, group, and genus/family level in 91%, 5%, and 2% of cases, respectively, in 20 min. In only 2% of these samples, MALDI-TOF MS did not yield any result. When blood cultures were multibacterial, identification was improved by using specific databases based on the Gram staining results. MALDI-TOF-MS is currently the fastest technique to accurately identify microorganisms grown in positive blood culture broths. PMID:20237092

  7. Aerobic Anoxygenic Phototrophic Bacteria

    PubMed Central

    Yurkov, Vladimir V.; Beatty, J. Thomas

    1998-01-01

    The aerobic anoxygenic phototrophic bacteria are a relatively recently discovered bacterial group. Although taxonomically and phylogenetically heterogeneous, these bacteria share the following distinguishing features: the presence of bacteriochlorophyll a incorporated into reaction center and light-harvesting complexes, low levels of the photosynthetic unit in cells, an abundance of carotenoids, a strong inhibition by light of bacteriochlorophyll synthesis, and the inability to grow photosynthetically under anaerobic conditions. Aerobic anoxygenic phototrophic bacteria are classified in two marine (Erythrobacter and Roseobacter) and six freshwater (Acidiphilium, Erythromicrobium, Erythromonas, Porphyrobacter, Roseococcus, and Sandaracinobacter) genera, which phylogenetically belong to the α-1, α-3, and α-4 subclasses of the class Proteobacteria. Despite this phylogenetic information, the evolution and ancestry of their photosynthetic properties are unclear. We discuss several current proposals for the evolutionary origin of aerobic phototrophic bacteria. The closest phylogenetic relatives of aerobic phototrophic bacteria include facultatively anaerobic purple nonsulfur phototrophic bacteria. Since these two bacterial groups share many properties, yet have significant differences, we compare and contrast their physiology, with an emphasis on morphology and photosynthetic and other metabolic processes. PMID:9729607

  8. Aerobic Dance in Public Schools.

    ERIC Educational Resources Information Center

    Chiles, Barbara Ann; Moore, Suzanne

    1981-01-01

    Aerobic dance offers a challenging workout in a social atmosphere. Though some physical education instructors tend to exclude dance units from the curriculum, most could teach aerobic dance if they had a basic knowledge of aerobic routines. The outline for a unit to be used in the class is presented. (JN)

  9. Managing for Improved Aerobic Stability

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aerobic deterioration or spoilage of silage is the result of aerobic microorganisms metabolizing components of the silage using oxygen. In the almost 40 years over which these silage conferences have been held, we have come to recognize the typical pattern of aerobic microbial development by which s...

  10. Aerobic and anaerobic cecal bacterial flora of commercially processed broilers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Differences in the bacterial flora of aerobic and anaerobic cultures of broiler ceca collected from a commercial poultry processing facility were determined. Bacterial isolates from cecal cultures were selected based on the ability of the bacteria to grow in media supplemented with lactate and succ...

  11. Flow microfluorometric analysis of phagocyte degranulation in bacteria-infected whole human blood cell cultures

    NASA Astrophysics Data System (ADS)

    Kravtsov, Alexander L.; Bobyleva, Elena V.; Grebenyukova, Tatyana P.; Kuznetsov, Oleg S.; Kulyash, Youri V.

    2002-07-01

    A quantitative flow microfluorometric method was used to study the intensity of human blood phagocyte degranulation in response to viable staphylococcus aureus or Yersinia pestis cells. Microorganisms were added directly to defibrinated whole blood. Uninfected and infected blood samples were incubated at 37 degrees C to 8 h. The results were recorded in dynamics after the staining of whole blood with acridine orange solution. Lymphocytes with a low azurophilic granule per cell content were discriminated from phagocytes by the measurement of single cell red cytoplasmic granule fluorescence. 30,000 cells in each sample were examined. S. aureus cells caused a dose-dependent decrease in the number of phagocytes having a high red cytoplasmic fluorescence intensity and a corresponding increase in the weakly fluorescence cell population. In the presence of an initial S. aureus-to-phagocyte ratio more than 1:1, degranulation was measured after 3 h of incubation and to 8 h the percentage of degranulated phagocytes was at least 100 percent Y. pestis cells grown for 48 h at 28 degrees C caused at same condition as the degranulation only about 50 percent of cells. Y.pestis EV cells preincubated in broth for 12 h at 37 degrees C did no stimulate the phahocyte degranulation. The results of these studies suggest that analysis of cell populations via flow microfluorimeter technology may be a powerful tool in analysis bacterial infection.

  12. Effectiveness of practices to reduce blood culture contamination: A Laboratory Medicine Best Practices systematic review and meta-analysis☆

    PubMed Central

    Snyder, Susan R.; Favoretto, Alessandra M.; Baetz, Rich Ann; Derzon, James H.; Madison, Bereneice M.; Mass, Diana; Shaw, Colleen S.; Layfield, Christopher D.; Christenson, Robert H.; Liebow, Edward B.

    2015-01-01

    Objectives This article is a systematic review of the effectiveness of three practices for reducing blood culture contamination rates: venipuncture, phlebotomy teams, and prepackaged preparation/collection (prep) kits. Design and methods The CDC-funded Laboratory Medicine Best Practices Initiative systematic review methods for quality improvement practices were used. Results Studies included as evidence were: 9 venipuncture (vs. versus intravenous catheter), 5 phlebotomy team; and 7 prep kit. All studies for venipuncture and phlebotomy teams favored these practices, with meta-analysis mean odds ratios for venipuncture of 2.69 and phlebotomy teams of 2.58. For prep kits 6 studies’ effect sizes were not statistically significantly different from no effect (meta-analysis mean odds ratio 1.12). Conclusions Venipuncture and the use of phlebotomy teams are effective practices for reducing blood culture contamination rates in diverse hospital settings and are recommended as evidence-based “best practices” with high overall strength of evidence and substantial effect size ratings. No recommendation is made for or against prep kits based on uncertain improvement. PMID:22709932

  13. Melatonin suppression of aerobic glycolysis (Warburg effect), survival signalling and metastasis in human leiomyosarcoma.

    PubMed

    Mao, Lulu; Dauchy, Robert T; Blask, David E; Dauchy, Erin M; Slakey, Lauren M; Brimer, Samantha; Yuan, Lin; Xiang, Shulin; Hauch, Adam; Smith, Kara; Frasch, Tripp; Belancio, Victoria P; Wren, Melissa A; Hill, Steven M

    2016-03-01

    Leiomyosarcoma (LMS) represents a highly malignant, rare soft tissue sarcoma with high rates of morbidity and mortality. Previously, we demonstrated that tissue-isolated human LMS xenografts perfused in situ are highly sensitive to the direct anticancer effects of physiological nocturnal blood levels of melatonin which inhibited tumour cell proliferative activity, linoleic acid (LA) uptake and metabolism to 13-hydroxyoctadecadienoic acid (13-HODE). Here, we show the effects of low pharmacological blood concentrations of melatonin following oral ingestion of a melatonin supplement by healthy adult human female subjects on tumour proliferative activity, aerobic glycolysis (Warburg effect) and LA metabolic signalling in tissue-isolated LMS xenografts perfused in situ with this blood. Melatonin markedly suppressed aerobic glycolysis and induced a complete inhibition of tumour LA uptake, 13-HODE release, as well as significant reductions in tumour cAMP levels, DNA content and [(3) H]-thymidine incorporation into DNA. Furthermore, melatonin completely suppressed the phospho-activation of ERK 1/2, AKT, GSK3β and NF-kB (p65). The addition of S20928, a nonselective melatonin antagonist, reversed these melatonin inhibitory effects. Moreover, in in vitro cell culture studies, physiological concentrations of melatonin repressed cell proliferation and cell invasion. These results demonstrate that nocturnal melatonin directly inhibited tumour growth and invasion of human LMS via suppression of the Warburg effect, LA uptake and other related signalling mechanisms. An understanding of these novel signalling pathway(s) and their association with aerobic glycolysis and LA metabolism in human LMS may lead to new circadian-based therapies for the prevention and treatment of LMS and potentially other mesenchymally derived solid tumours. PMID:26607298

  14. Rapid Identification of Bacteria from Positive Blood Cultures by Fluorescence-Based PCR–Single-Strand Conformation Polymorphism Analysis of the 16S rRNA Gene

    PubMed Central

    Turenne, Christine Y.; Witwicki, Evelyn; Hoban, Daryl J.; Karlowsky, James A.; Kabani, Amin M.

    2000-01-01

    Bacteremia continues to result in significant morbidity and mortality, particularly in patients who are immunocompromised. Currently, patients with suspected bacteremia are empirically administered broad-spectrum antibiotics, as definitive diagnosis relies upon the use of blood cultures, which impose significant delays in and limitations to pathogen identification. To address the limitations of growth-based identification, the sequence variability of the 16S rRNA gene of bacteria was targeted for rapid identification of bacterial pathogens isolated directly from blood cultures using a fluorescence-based PCR–single-strand conformation polymorphism (SSCP) protocol. Species-specific SSCP patterns were determined for 25 of the most common bacterial species isolated from blood cultures; these isolates subsequently served as a reference collection for bacterial identification for new cases of bacteremia. A total of 272 blood-culture-positive patient specimens containing bacteria were tested. A previously determined SSCP pattern was observed for 251 (92%) specimens, with 21 (8%) specimens demonstrating SSCP patterns distinct from those in the reference collection. Time to identification from blood culture positivity ranged from 1 to 8 days with biochemical testing, whereas identification by fluorescence-based capillary electrophoresis was obtained as early as 7 h at a calculated cost of $10 (U.S. currency) per specimen when tested in batches of 10. Limitations encountered included the inability to consistently detect mixed cultures as well as some species demonstrating identical SSCP patterns. This method can be applied directly to blood cultures or whole-blood specimens, where early pathogen identification would result in a timely diagnosis with possible implications for patient management costs and the mortality and morbidity of infections. PMID:10655337

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

    PubMed

    Gharbi, M; Latrach, R; Sassi, L; Darghouth, M A

    2012-08-01

    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. PMID:22910672

  16. Method for Radiorespirometric Detection of Bacteria in Pure Culture and in Blood

    PubMed Central

    Schrot, J. Rudolph; Hess, Walter C.; Levin, Gilbert V.

    1973-01-01

    Methods are described for the detection of low numbers of bacteria by monitoring 14CO2 evolved from 14C-labeled substrates. Cell suspensions are filtered with membrane filters, and the filter is then moistened with 0.1 ml of labeled medium in a small, closed apparatus. Evolved 14CO2 is collected with Ba(OH)2-moistened filter pads and assayed with conventional radioactivity counting equipment. The kinetics of 14CO2 evolution are shown for several species of bacteria. Fewer than 100 colony-forming units of most species tested were detected in 2 h or less. Bacteria were inoculated into blood and the mixture was treated to lyse the blood cells. The suspension ws filtered and the filter was placed in a small volume of labeled medium. The evolved 14CO2 was trapped and counted. A key development in the methodology was finding that an aqueous solution of Rhyozyme and Triton X-100 produced lysis of blood but was not detrimental to bacteria. PMID:4588194

  17. Histamine Induces Alzheimer's Disease-Like Blood Brain Barrier Breach and Local Cellular Responses in Mouse Brain Organotypic Cultures

    PubMed Central

    Sedeyn, Jonathan C.; Wu, Hao; Hobbs, Reilly D.; Levin, Eli C.; Nagele, Robert G.; Venkataraman, Venkat

    2015-01-01

    Among the top ten causes of death in the United States, Alzheimer's disease (AD) is the only one that cannot be cured, prevented, or even slowed down at present. Significant efforts have been exerted in generating model systems to delineate the mechanism as well as establishing platforms for drug screening. In this study, a promising candidate model utilizing primary mouse brain organotypic (MBO) cultures is reported. For the first time, we have demonstrated that the MBO cultures exhibit increased blood brain barrier (BBB) permeability as shown by IgG leakage into the brain parenchyma, astrocyte activation as evidenced by increased expression of glial fibrillary acidic protein (GFAP), and neuronal damage-response as suggested by increased vimentin-positive neurons occur upon histamine treatment. Identical responses—a breakdown of the BBB, astrocyte activation, and neuronal expression of vimentin—were then demonstrated in brains from AD patients compared to age-matched controls, consistent with other reports. Thus, the histamine-treated MBO culture system may provide a valuable tool in combating AD. PMID:26697497

  18. Histamine Induces Alzheimer's Disease-Like Blood Brain Barrier Breach and Local Cellular Responses in Mouse Brain Organotypic Cultures.

    PubMed

    Sedeyn, Jonathan C; Wu, Hao; Hobbs, Reilly D; Levin, Eli C; Nagele, Robert G; Venkataraman, Venkat

    2015-01-01

    Among the top ten causes of death in the United States, Alzheimer's disease (AD) is the only one that cannot be cured, prevented, or even slowed down at present. Significant efforts have been exerted in generating model systems to delineate the mechanism as well as establishing platforms for drug screening. In this study, a promising candidate model utilizing primary mouse brain organotypic (MBO) cultures is reported. For the first time, we have demonstrated that the MBO cultures exhibit increased blood brain barrier (BBB) permeability as shown by IgG leakage into the brain parenchyma, astrocyte activation as evidenced by increased expression of glial fibrillary acidic protein (GFAP), and neuronal damage-response as suggested by increased vimentin-positive neurons occur upon histamine treatment. Identical responses-a breakdown of the BBB, astrocyte activation, and neuronal expression of vimentin-were then demonstrated in brains from AD patients compared to age-matched controls, consistent with other reports. Thus, the histamine-treated MBO culture system may provide a valuable tool in combating AD. PMID:26697497

  19. Bacteriological Profile and Antimicrobial Susceptibility Pattern of Blood Culture Isolates among Septicemia Suspected Children in Selected Hospitals Addis Ababa, Ethiopia

    PubMed Central

    Negussie, Adugna; Mulugeta, Gebru; Bedru, Ahmed; Ali, Ibrahim; Shimeles, Damte; Lema, Tsehaynesh; Aseffa, Abraham

    2015-01-01

    Background Blood stream infections are major cause of morbidity and mortality in children in developing countries. The emerging of causative agents and resistance to various antimicrobial agents are increased from time to time. The main aim of this study was to determine the bacterial agents and antimicrobial susceptibility patterns among children suspected of having septicemia. Methods A cross sectional study involved about 201 pediatric patients (≤ 12 years) was conducted from October 2011 to February 2012 at pediatric units of TikurAnbessa Specialized Hospital and Yekatit 12 Hospital. Standard procedure was followed for blood sample collection, isolate identifications and antimicrobial susceptibility testing. Results Among 201 study subjects 110 (54.7%) were males. Majority 147 (73.1%) of them were neonates (≤ 28 days). The mean length of hospital stay before sampling was 4.29 days. Out of the 201 tested blood samples, blood cultures were positive in 56 (27.9%).Gram negative and Gram positive bacteria constituted 29(51.8%) and 26(46.4%), respectively. The most frequent pathogen found was Staphylococcus aureus 13 (23.2%), followed by Serratia marcescens 12(21.4%), CoNS 11(19.6%), klebsiella spp 9(16%) and Salmonella spp 3(5.4%). Majority of bacterial isolates showed high resistance to Ampicillin, Penicillin, Co-trimoxazole, Gentamicin and Tetracycline which commonly used in the study area. Conclusion Majority of the isolates were multidrug resistant. These higher percentages of multi-drug resistant emerged isolates urge us to take infection prevention measures and to conduct other large studies for appropriate empiric antibiotic choice. PMID:26997847

  20. Characterization of Nasal and Blood Culture Isolates of Methicillin-Resistant Staphylococcus aureus from Patients in United States Hospitals

    PubMed Central

    Tickler, Isabella A.; Goering, Richard V.; Kreiswirth, Barry N.; Mediavilla, José R.; Persing, David H.

    2012-01-01

    A total of 299 nares and 194 blood isolates of methicillin-resistant Staphylococcus aureus (MRSA), each recovered from a unique patient, were collected from 23 U.S. hospitals from May 2009 to March 2010. All isolates underwent spa and staphylococcal cassette chromosome mec element (SCCmec) typing and antimicrobial susceptibility testing; a subset of 84 isolates was typed by pulsed-field gel electrophoresis (PFGE) using SmaI. Seventy-six spa types were observed among the isolates. Overall, for nasal isolates, spa type t002-SCCmec type II (USA100) was the most common strain type (37% of isolates), while among blood isolates, spa type t008-SCCmec type IV (USA300) was the most common (39%). However, the proportion of all USA100 and USA300 isolates varied by United States census region. Nasal isolates were more resistant to tobramycin and clindamycin than blood isolates (55.9% and 48.8% of isolates versus 36.6% and 39.7%, respectively; for both, P < 0.05). The USA300 isolates were largely resistant to fluoroquinolones. High-level mupirocin resistance was low among all spa types (<5%). SCCmec types III and VIII, which are rare in the United States, were observed along with several unusual PFGE types, including CMRSA9, EMRSA15, and the PFGE profile associated with sequence type 239 (ST239) isolates. Typing data from this convenience sample suggest that in U.S. hospitalized patients, USA100 isolates of multiple spa types, while still common in the nares, have been replaced by USA300 isolates as the predominant MRSA strain type in positive blood cultures. PMID:22155818

  1. Development of o.a.s.i.s., a new automated blood culture system in which detection is based on measurement of bottle headspace pressure changes.

    PubMed Central

    Stevens, C M; Swaine, D; Butler, C; Carr, A H; Weightman, A; Catchpole, C R; Healing, D E; Elliott, T S

    1994-01-01

    o.a.s.i.s. (Unipath Ltd., Basingstoke, United Kingdom) is a new automated blood culture system. The metabolism of microorganisms is detected by measuring changes in the pressure of the headspace of blood culture bottles. These changes are measured by monitoring the position of a flexible sealing septum, every 5 min, with a scanning laser sensor. This noninvasive system can detect both gas absorption and production and does not rely solely on measuring increasing carbon dioxide levels. A research prototype instrument was used to carry out an evaluation of the media, the detection system, and its associated detection algorithm. In simulated blood cultures, o.a.s.i.s. supported growth and detected a range of clinical isolates. Times to positivity were significantly shorter in o.a.s.i.s. than in the BACTEC 460 system. Results of a clinical feasibility study, with a manual blood culture system as a control, confirmed that o.a.s.i.s. was able to support the growth and detection of a variety of clinically significant organisms. On the basis of these findings, full-scale comparative clinical trials of o.a.s.i.s. with other automated blood culture systems are warranted. PMID:7929769

  2. Aerobic microbial mineralization of dichloroethene as sole carbon substrate

    USGS Publications Warehouse

    Bradley, P.M.; Chapelle, F.H.

    2000-01-01

    Microorganisms indigenous to the bed sediments of a black- water stream utilized 1,2-dichloroethene (1,2-DCE) as a sole carbon substrate for aerobic metabolism. Although no evidence of growth was observed in the minimal salts culture media used in this study, efficient aerobic microbial mineralization of 1,2-DCE as sole carbon substrate was maintained through three sequential transfers (107 final dilution) of the original environmental innoculum. These results indicate that 1,2-DCE can be utilized as a primary substrate to support microbial metabolism under aerobic conditions.Microorganisms indigenous to the bed sediments of a black-water stream utilized 1,2-dichloroethene (1,2-DCE) as a sole carbon substrate for aerobic metabolism. Although no evidence of growth was observed in the minimal salts culture media used in this study, efficient aerobic microbial mineralization of 1,2-DCE as sole carbon substrate was maintained through three sequential transfers (107 final dilution) of the original environmental innoculum. These results indicate that 1,2-DCE can be utilized as a primary substrate to support microbial metabolism under aerobic conditions.

  3. Kinetic and inhibition studies for the aerobic cometabolism of 1,1,1-trichloroethane, 1,1-dichloroethylene, and 1,1-dichloroethane by a butane-grown mixed culture.

    PubMed

    Kim, Young; Arp, Daniel J; Semprini, Lewis

    2002-12-01

    Batch kinetic and inhibition studies were performed for the aerobic cometabolism of 1,1,1-trichloroethane (1,1,1-TCA), 1,1-dichloroethylene (1,1-DCE), and 1,1-dichloroethane (1,1-DCA) by a butane-grown mixed culture. These chlorinated aliphatic hydrocarbons (CAHs) are often found together as cocontaminants in groundwater. The maximum degradation rates (k(max)) and half-saturation coefficients (K(s)) were determined in single compound kinetic tests. The highest k(max) was obtained for butane (2.6 micromol/mg TSS/h) followed by 1,1-DCE (1.3 micromol/mg TSS/h), 1,1-DCA (0.49 micromol/mg TSS/h), and 1,1,1-TCA (0.19 micromol/mg TSS/h), while the order of K(s) from the highest to lowest was 1,1-DCA (19 microM), butane (19 microM), 1,1,1-TCA (12 microM) and 1,1-DCE (1.5 microM). The inhibition types were determined using direct linear plots, while inhibition coefficients (K(ic) and K(iu)) were estimated by nonlinear least squares regression (NLSR) fits to the kinetic model of the identified inhibition type. Two different inhibition types were observed among the compounds. Competitive inhibition among CAHs was indicated from direct linear plots, and the CAHs also competitively inhibited butane utilization. 1,1-DCE was a stronger inhibitor than the other CAHs. Mixed inhibition of 1,1,1-TCA, 1,1-DCA, and 1,1-DCE transformations by butane was observed. Thus, both competitive and mixed inhibitions are important in cometabolism of CAHs by this butane culture. For competitive inhibition between CAHs, the ratio of the K(s) values was a reasonable indicator of competitive inhibition observed. Butane was a strong inhibitor of CAH transformation, having a much lower inhibition coefficient than the K(s) value of butane, while the CAHs were weak inhibitors of butane utilization. Model simulations of reactor systems where both the growth substrate and the CAHs are present indicate that reactor performance is significantly affected by inhibition type and inhibition coefficients. Thus

  4. Degradation of TCE using sequential anaerobic biofilm and aerobic immobilized bed reactor

    NASA Technical Reports Server (NTRS)

    Chapatwala, Kirit D.; Babu, G. R. V.; Baresi, Larry; Trunzo, Richard M.

    1995-01-01

    Bacteria capable of degrading trichloroethylene (TCE) were isolated from contaminated wastewaters and soil sites. The aerobic cultures were identified as Pseudomonas aeruginosa (four species) and Pseudomonas fluorescens. The optimal conditions for the growth of aerobic cultures were determined. The minimal inhibitory concentration values of TCE for Pseudomonas sps. were also determined. The aerobic cells were immobilized in calcium alginate in the form of beads. Degradation of TCE by the anaerobic and dichloroethylene (DCE) by aerobic cultures was studied using dual reactors - anaerobic biofilm and aerobic immobilized bed reactor. The minimal mineral salt (MMS) medium saturated with TCE was pumped at the rate of 1 ml per hour into the anaerobic reactor. The MMS medium saturated with DCE and supplemented with xylenes and toluene (3 ppm each) was pumped at the rate of 1 ml per hour into the fluidized air-uplift-type reactor containing the immobilized aerobic cells. The concentrations of TCE and DCE and the metabolites formed during their degradation by the anaerobic and aerobic cultures were monitored by GC. The preliminary study suggests that the anaerobic and aerobic cultures of our isolates can degrade TCE and DCE.

  5. 16S rRNA Gene Sequence-Based Identification of Bacteria in Automatically Incubated Blood Culture Materials from Tropical Sub-Saharan Africa

    PubMed Central

    Schwarz, Norbert Georg; Hahn, Andreas; Boahen, Kennedy; Sarpong, Nimako; Adu-Sarkodie, Yaw; Halbgewachs, Eva; Marks, Florian; von Kalckreuth, Vera; Poppert, Sven; Loderstaedt, Ulrike; May, Jürgen; Hagen, Ralf Matthias

    2015-01-01

    Background 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. Methods 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. Results 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. Conclusions 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. PMID:26270631

  6. An epidemiological study of blood culture isolates of coagulase-negative staphylococci demonstrating hospital-acquired infection.

    PubMed Central

    Burnie, J P; Naderi-Nasab, M; Loudon, K W; Matthews, R C

    1997-01-01

    We applied pulsed-field gel electrophoresis (PFGE) after SmaI digestion and random amplification of polymorphic DNA (RAPD) analysis with nine oligonucleotide primers to 146 blood culture isolates of Staphylococcus epidermidis and 25 blood culture isolates of Staphylococcus haemolyticus. These were obtained over a 12-month period from patients on the neonatal and hematology units of the Central Manchester Health Care Trust. PFGE demonstrated two clusters of isolates of S. epidermidis (type A and type B) on the neonatal ward and a single cluster (type C) on the hematology unit. Type A was represented by 10 indistinguishable isolates from nine patients, type B was represented by 20 isolates from 14 patients, and type C was represented by 26 isolates from 10 patients. Type A isolates were resistant to chloramphenicol and type C isolates were resistant to ciprofloxacin, mirroring current antibiotic usage. There was no evidence of cross infection due to S. haemolyticus. RAPD analysis, on the basis of a single band difference, produced 58 types of S. epidermidis and 12 types of S. haemolyticus with primer 8 (ATG TAA GCT CCT GGG GAT TCA C; 5' to 3') and 54 types of S. epidermidis and 10 types of S. haemolyticus with primer 9 (AAG TAA GTG ACT GGG GTG AGC G; 5' to 3'). Combining the results confirmed cross infection. Types A, B, and C were concurrently isolated from the hands of the staff of the appropriate unit. Partial control was achieved by withdrawing ciprofloxacin use in the case of the hematology unit and improving hand hygiene in both units. PMID:9196185

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

    PubMed

    Rahiman, F; Pool, E J

    2014-01-01

    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

  8. Comparative Study in Early Neonates with Septicemia by Blood Culture, Staining Techniques and C – Reactive Protein (CRP)

    PubMed Central

    Dhanalakshmi, V.

    2015-01-01

    Aim: The aim of this study was to compare and evaluate the pathogenic bacteria in neo-natal septicemia by using various diagnostic techniques. Setting and Design: Our study was designed to evaluate a feasible method to diagnose neonatal septicemia even at primary health centre level. Materials and Methods: Blood samples were collected aseptically from 70 neonates. The specimens were inoculated into brain heart infusion broth and subcultures were performed with specific media. Antibiotic sensitivity pattern of isolates was studied by Modified Kirby Bauer Disc diffusion technique and differentiate the isolates by staining methods. C-reactive protein (CRP) was evaluated by using standard kit method. Results: Out of 70 cases of childhood septicemia of age group 1-30 days, 37 had positive CRP, 36 were positive for BCS and blood culture was positive only in 41 cases, where predominant organism being Klebsiella species (n=28, 68.29%) followed by Escherichia coli (n=4, 9.76%), Pseudomonas aeruginosa (n=3,7.31%), Proteus mirabilis (n=2,4.88%) and Coagulase negative staphylococcus (n=4,9.76%). Conclusion: Our findings suggest that Klebsiella species as an important cause of neonatal septicemia. The isolated organisms were found to be highly sensitive to cefatoxime and amikacin. Hence, these antibiotics can be considered as the first drug of choice for neonatal septicemia. PMID:25954618

  9. Evaluation of an Automated Rapid Diagnostic Assay for Detection of Gram-Negative Bacteria and Their Drug-Resistance Genes in Positive Blood Cultures

    PubMed Central

    Tojo, Masayoshi; Fujita, Takahiro; Ainoda, Yusuke; Nagamatsu, Maki; Hayakawa, Kayoko; Mezaki, Kazuhisa; Sakurai, Aki; Masui, Yoshinori; Yazaki, Hirohisa; Takahashi, Hiroshi; Miyoshi-Akiyama, Tohru; Totsuka, Kyoichi; Kirikae, Teruo; Ohmagari, Norio

    2014-01-01

    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 blaCTX-M, 119 blaIMP, 8 blaKPC, 16 blaNDM, 24 blaOXA-23, 1 blaOXA-24/40, 1 blaOXA-48, 4 blaOXA-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. PMID:24705449

  10. L-arginine and arginine analogues: effects on isolated blood vessels and cultured endothelial cells.

    PubMed Central

    Schmidt, H. H.; Baeblich, S. E.; Zernikow, B. C.; Klein, M. M.; Böhme, E.

    1990-01-01

    1. The present study examined effects of arginine (Arg) and various Arg analogues on the vascular tone of rabbit and rat aortic rings, the release of nitrite from cultured bovine aortic endothelial cells and the metabolism of L-Arg in bovine and porcine endothelial cell homogenates. The respective D-enantiomers or N-alpha-benzoyl-L-arginine ethyl ester did not substitute for L-Arg. 2. In bovine aortic endothelial cells, the release of nitrite was only observed in the presence of L-Arg or L-Arg methyl ester in the cell culture medium. 3. In dialyzed homogenates of porcine and bovine aortic endothelial cells, L-Arg was metabolized independently of NADPH and Ca2+ to yield L-ornithine (L-Orn) and L-citrulline (L-Cit). No concomitant nitrite formation was detected. 4. Pretreatment of rabbit and rat aortic rings with L-canavanine (L-Can) or NG-monomethyl-L-Arg (L-NMMA) inhibited ATP- and acetylcholine-induced relaxations (endothelium-dependent) but not glyceryltrinitrate-induced relaxations (endothelium-independent). 5. In rabbit aortic rings, Arg and monomeric Arg analogues induced endothelium-independent relaxations. L-Arg methyl ester induced an endothelium-independent contraction, and L-NMMA induced a relaxation in the absence of endothelium and a contraction in the presence of endothelium. Polymeric basic amino acids such as poly L-Arg induced endothelium-dependent relaxations (inhibited by L-Can), a subsequent refractoriness to endothelium-dependent vasodilators (not prevented by L-Can) and endothelial cell death.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2282457

  11. Aerobic landfill bioreactor

    DOEpatents

    Hudgins, Mark P; Bessette, Bernard J; March, John C; McComb, Scott T.

    2002-01-01

    The present invention includes a system of decomposing municipal solid waste (MSW) within a landfill by converting the landfill to aerobic degradation in the following manner: (1) injecting air via the landfill leachate collection system (2) injecting air via vertical air injection wells installed within the waste mass; (3) applying leachate to the waste mass using a pressurized drip irrigation system; (4) allowing landfill gases to vent; and (5) adjusting air injection and recirculated leachate to achieve a 40% to 60% moisture level and a temperature between 120.degree. F. and 140.degree. F. in steady state.

  12. Aerobic landfill bioreactor

    DOEpatents

    Hudgins, Mark P; Bessette, Bernard J; March, John; McComb, Scott T.

    2000-01-01

    The present invention includes a method of decomposing municipal solid waste (MSW) within a landfill by converting the landfill to aerobic degradation in the following manner: (1) injecting air via the landfill leachate collection system (2) injecting air via vertical air injection wells installed within the waste mass; (3) applying leachate to the waste mass using a pressurized drip irrigation system; (4) allowing landfill gases to vent; and (5) adjusting air injection and recirculated leachate to achieve a 40% to 60% moisture level and a temperature between 120.degree. F. and 140.degree. F. in steady state.

  13. [Blood cultures in the paediatric emergency department. Guidelines and recommendations on their indications, collection, processing and interpretation].

    PubMed

    Hernández-Bou, S; Álvarez Álvarez, C; Campo Fernández, M N; García Herrero, M A; Gené Giralt, A; Giménez Pérez, M; Piñeiro Pérez, R; Gómez Cortés, B; Velasco, R; Menasalvas Ruiz, A I; García García, J J; Rodrigo Gonzalo de Liria, C

    2016-05-01

    Blood culture (BC) is the gold standard when a bacteraemia is suspected, and is one of the most requested microbiological tests in paediatrics. Some changes have occurred in recent years: the introduction of new vaccines, the increasing number of patients with central vascular catheters, as well as the introduction of continuous monitoring BC systems. These changes have led to the review and update of different factors related to this technique in order to optimise its use. A practice guideline is presented with recommendations on BC, established by the Spanish Society of Paediatric Emergency Care and the Spanish Society for Paediatric Infectious Diseases. After reviewing the available scientific evidence, several recommendations for each of the following aspects are presented: BC indications in the Emergency Department, how to obtain, transport and process cultures, special situations (indications and interpretation of results in immunosuppressed patients and/or central vascular catheter carriers, indications for anaerobic BC), differentiation between bacteraemia and contamination when a BC shows bacterial growth and actions to take with a positive BC in patients with fever of unknown origin. PMID:26227314

  14. Rapid identification of Acinetobacter spp. by fluorescence in situ hybridization (FISH) from colony and blood culture material

    PubMed Central

    Essig, A.; Hagen, R. M.; Riecker, M.; Jerke, K.; Ellison, D.; Poppert, S.

    2011-01-01

    Multi-drug-resistant strains of the Acinetobacter baumannii complex cause nosocomial infections. Rapid identification of Acinetobacter spp. is desirable in order to facilitate therapeutic or hygiene decisions. We evaluated a newly designed DNA probe that can be used under standard conditions in both a microwave oven and a slide chamber for the rapid identification of Acinetobacter spp. by fluorescence in situ hybridization (FISH). Using FISH, the new probe correctly identified 81/81 Acinetobacter spp. isolates and excluded 109/109 tested non-target organisms from agar culture. Furthermore, the new probe correctly identified 7/7 Acinetobacter spp. in 214 blood cultures determined to contain Gram-negative bacteria by Gram staining. Using either the microwave oven or slide chamber technique, the new probe was able to identify Acinetobacter spp. in 100% of the samples tested. FISH used in conjunction with our newly designed probe provides an easy, cheap, precise, and rapid method for the preliminary identification of Acinetobacter spp., especially in laboratories where more sophisticated methods like matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) are not available. PMID:24516735

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

    NASA Technical Reports Server (NTRS)

    Lawless, DeSales

    2003-01-01

    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.

  16. Aerobic Fitness for the Severely and Profoundly Mentally Retarded.

    ERIC Educational Resources Information Center

    Bauer, Dan

    1981-01-01

    The booklet discusses the aerobic fitness capacities of severely/profoundly retarded students and discusses approaches for improving their fitness. An initial section describes a method for determining the student's present fitness level on the basis of computations of height, weight, blood pressure, resting pulse, and Barach Index and Crampton…

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

    PubMed

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

    2012-08-30

    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

  18. Characterization of aerobic ethanol productions in a computerized auxostat

    SciTech Connect

    Fraleigh, S.P.

    1989-01-01

    For many valuable bioproducts high productivity is associated with rapid growth. However, most continuous microbial cultures become unstable when the dilution rate is fixed near the value for maximum growth rate. The auxostat culture technique employs feedback control of a nutrient or metabolite to stabilize the biomass at its maximum potential growth rate. An auxostat device is therefore ideal for study of bioprocesses involving the overproduction of primary metabolites such as ethanol. Oxidoreductive transformations involving ethanol are utilized by Saccharomyces yeasts when normal respiration cannot satisfy energy needs. When rapid growth or other stress creates oxidoreductive conditions in aerobic Saccharomyces cultures, very high specific ethanol formation rates are established and biomass yield drops to levels more typical of anaerobic fermentation. Although the physiology is favorable, the potential for large-scale aerobic ethanol processes to compete with traditional anaerobic fermentations has not previously been assessed. In this study, a fully computerized auxostat device was constructed and used to characterize the specific and volumetric aerobic ethanol productivity of the yeast Saccharomyces cerevisiae. To divert substrate away from biomass and into product formation, aerobic cultures were stressed with variations of ionic balance (via extreme K{sup +} and H{sup +} setpoints) in the auxostat device. During growth with limiting K{sup +} concentrations, the goal of very low biomass yield was attained but the rate of ethanol production was poor. However, with excess K{sup +} the volumetric productivity reached 6.1 g/I,-h, a value that is comparable to optimized, continuous anaerobic cultures.

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

    PubMed Central

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

    2016-01-01

    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

  20. Aerobic Fitness and School Children.

    ERIC Educational Resources Information Center

    Hinkle, J. Scott

    1997-01-01

    Provides school counselors with information on aerobic exercise (specifically running) and the psychological, behavioral, and physical benefits children obtained by participating in fitness programs. Recommends collaboration between school counselors and physical education teachers and gives a preliminary discussion of aerobic running and its…

  1. Aerobic Fitness and School Children.

    ERIC Educational Resources Information Center

    Hinkle, J. Scott

    1992-01-01

    Provides school counselors with information regarding aerobic exercise (specifically running), and the psychological, behavioral, and physical benefits children obtain by participating in fitness programs. Presents methods of collaboration between school counselors and physical education teachers. Offers preliminary discussion of aerobic running…

  2. Exercise, Animal Aerobics, and Interpretation?

    ERIC Educational Resources Information Center

    Oliver, Valerie

    1996-01-01

    Describes an aerobic activity set to music for children that mimics animal movements. Example exercises include walking like a penguin or jumping like a cricket. Stresses basic aerobic principles and designing the program at the level of children's motor skills. Benefits include reaching people who normally don't visit nature centers, and bridging…

  3. Uterine culture in mares.

    PubMed

    Brook, D

    1984-05-01

    A guarded, sterile swab is used to obtain samples for uterine culture. With the mare in stocks, the tail bandage and the perineum washed, the culture rod is introduced into the vagina with a gloved hand. After the rod is guided through the cervix, the guard cap is dislodged and the swab is rubbed along the endometrium, after which the rod is extracted. Samples for uterine culture should only be obtained during full estrus. Swabs should be directly plated onto agar within 2 hours of collection. Blood agar is appropriate for initial screening, but use of specialized types of agar expedites identification of microbes. Plates are incubated at 37 C and inspected for growth every 12 hours. The type and number of bacterial colonies should be coupled with the history and clinical signs in deciding on the necessity and type of treatment. Pure, heavy bacterial growth is usually accompanied by clinical signs of infection. Interpretation of the significance of moderate bacterial growth may be aided by cytologic examination of endometrial smears, made by rolling the swab onto a glass slide and staining with Diff - Quik . Large numbers of neutrophils indicate the need for antibiotic therapy. Mixed bacterial growth and variable numbers of neutrophils usually indicate faulty sampling technic. Microaerophilic or anaerobic cultures may aid diagnosis in cases of equivocal aerobic culture results. PMID:6377040

  4. Genome Sequence of a Virulent Pseudomonas aeruginosa Strain, 12-4-4(59), Isolated from the Blood Culture of a Burn Patient

    PubMed Central

    Karna, S. L. Rajasekhar; Chen, Tsute; Chen, Ping; Peacock, Trent J.; Abercrombie, Johnathan J.

    2016-01-01

    Pseudomonas aeruginosa is an opportunistic pathogen that frequently infects wounds, significantly impairs wound healing, and causes morbidity and mortality in burn patients. Here, we report the genome sequence of a virulent strain of P. aeruginosa, 12-4-4(59), isolated from the blood culture of a burn patient. PMID:26941150

  5. EFFECTS OF PRE-INCUBATION HOLDING TIME AND TEMPERATURE ON INTERFERON-GAMMA RESPONSES IN WHOLE BLOOD CULTURES FROM MYCOBACTERIUM BOVIS-INFECTED CATTLE

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The BovigamTM assay is approved for use within the U.S. as a complementary diagnostic test for tuberculosis. The in vitro assay detects interferon (IFN)-gamma produced in whole blood cultures stimulated with specific antigen. Stimulants commonly used in the assay are purified protein derivatives d...

  6. Aerobic biodegradation process of petroleum and pathway of main compounds in water flooding well of Dagang oil field.

    PubMed

    Cai, Minmin; Yao, Jun; Yang, Huaijun; Wang, Ruixia; Masakorala, Kanaji

    2013-09-01

    Aerobic biodegradation of crude oil and its pathways were investigated via in vitro culture and GC-MS analysis in water flooding wells of Dagang oil field. The in vitro aerobic culture lasted 90 days when 99.0% of n-alkanes and 43.03-99.9% of PAHs were degraded and the biomarkers and their ratios were changed. The spectra of components in the residual oil showed the similar biodegradation between aerobic process of 90 days and degradation in reservoir which may last for some millions years, and the potential of serious aerobic biodegradation of petroleum in reservoir. 24 Metabolites compounds were separated and identified from aerobic culture, including fatty acid, naphthenic acid, aromatic carboxylic acid, unsaturated acid, alcohols, ketones and aldehydes. The pathways of alkanes and aromatics were proposed, which suggests that oxidation of hydrocarbon to organic acid is an important process in the aerobic biodegradation of petroleum. PMID:23867530

  7. Preparation of Glycerol-Enriched Yeast Culture and Its Effect on Blood Metabolites and Ruminal Fermentation in Goats

    PubMed Central

    Ye, Gengping; Zhu, Yongxing; Liu, Jin; Chen, Xingxiang; Huang, Kehe

    2014-01-01

    The aim of this study was to isolate a glycerol-producing yeast strain from nature to prepare glycerol-enriched yeast culture (GY), and preliminarily evaluate the effects of GY on blood metabolites and ruminal fermentation in goats. During the trial, six isolates were isolated from unprocessed honey, and only two isolates with higher glycerol yield were identified by analysis of 26S ribosomal DNA sequences. One of the two isolates was identified as Saccharomyces cerevisiae, a direct-fed microbe permitted by the FDA. This isolate was used to prepare GY. The fermentation parameters were optimized through single-factor and orthogonal design methods to maximize the glycerol yield and biomass. The final GY contained 38.7±0.6 g/L glycerol and 12.6±0.5 g/L biomass. In vivo, eight castrated male goats with ruminal fistula were used in a replicated 4×4 Latin square experiment with four consecutive periods of 15 d. Treatments were as follows: control, LGY, MGY, and HGY with 0, 100, 200, and 300 mL GY per goat per day, respectively. The GY was added in two equal portions at 08∶00 and 17∶00 through ruminal fistula. Samples of blood and ruminal fluid were collected on the last one and two days of each period, respectively. Results showed that the plasma concentrations of triglyceride and total cholesterol were not affected by the supplemented GY. Compared with the control, goats supplemented with MGY and HGY had significantly higher (P<0.05) concentrations of plasma glucose and total protein, ruminal volatile fatty acid and molar proportion of propionate, and significantly lower (P<0.05) ruminal pH and ammonia nitrogen. These parameters changed linearly with increasing GY supplementation level (P<0.05). In conclusion, GY has great potential to be developed as a feed additive with dual effects of glycerol and yeast for ruminants. PMID:24709881

  8. In vitro models of the blood-brain barrier: An overview of commonly used brain endothelial cell culture models and guidelines for their use.

    PubMed

    Helms, Hans C; Abbott, N Joan; Burek, Malgorzata; Cecchelli, Romeo; Couraud, Pierre-Olivier; Deli, Maria A; Förster, Carola; Galla, Hans J; Romero, Ignacio A; Shusta, Eric V; Stebbins, Matthew J; Vandenhaute, Elodie; Weksler, Babette; Brodin, Birger

    2016-05-01

    The endothelial cells lining the brain capillaries separate the blood from the brain parenchyma. The endothelial monolayer of the brain capillaries serves both as a crucial interface for exchange of nutrients, gases, and metabolites between blood and brain, and as a barrier for neurotoxic components of plasma and xenobiotics. This "blood-brain barrier" function is a major hindrance for drug uptake into the brain parenchyma. Cell culture models, based on either primary cells or immortalized brain endothelial cell lines, have been developed, in order to facilitate in vitro studies of drug transport to the brain and studies of endothelial cell biology and pathophysiology. In this review, we aim to give an overview of established in vitro blood-brain barrier models with a focus on their validation regarding a set of well-established blood-brain barrier characteristics. As an ideal cell culture model of the blood-brain barrier is yet to be developed, we also aim to give an overview of the advantages and drawbacks of the different models described. PMID:26868179

  9. Growth of Campylobacter Incubated Aerobically in Media Supplemented with Peptones

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Growth of Campylobacter cultures incubated aerobically in media supplemented with peptones was studied, and additional experiments were conducted to compare growth of the bacteria in media supplemented with peptones to growth in media supplemented with fumarate-pyruvate-minerals-vitamins (FPMV). A b...

  10. Culturally-sensitive weight loss program produces significant reduction in weight, blood pressure, and cholesterol in eight weeks.

    PubMed Central

    Ard, J. D.; Rosati, R.; Oddone, E. Z.

    2000-01-01

    Dietary and behavioral needs of special populations are rarely considered in traditional weight loss programs. This study assessed the impact of culturally-sensitive modifications to the Duke University Rice Diet weight loss program for African-American dieters. The study was a randomized modified cross-over study in which volunteers received either early or delayed weight loss intervention. Final outcomes were measured at 8 weeks. At the onset of the study, there were 56 African American participants, however, only 44 (79%) completed the study. The eight-week intervention was a modified 1000-calorie/day version of the Rice Diet. Modifications to the program included decreased cost, culturally-sensitive recipes, addressing attitudes about exercise, and including family members in weight loss efforts. Average weight loss for subjects completing the program was 14.8 pounds (SD = 6.8 pounds). BMI decreased from 37.8 kg/m2 to 35.3 kg/m2 (p < 0.01). Total cholesterol levels decreased from 199.2 mg/dL to 185.4 mg/dL (p < 0.01); systolic and diastolic blood pressure decreased by 4.3 mmHg (p < 0.01) and 2.4 mmHg (p < 0.05), respectively. The control group showed no significant change in any outcome measures. We found that diet programs can be successfully tailored to incorporate the needs of African-Americans. Most importantly, these dietary program changes can lead to significant improvement in clinical parameters. Additional studies are necessary to determine the permanence of these short-term changes. PMID:11152083

  11. Cyclosporine A kinetics in brain cell cultures and its potential of crossing the blood-brain barrier.

    PubMed

    Bellwon, P; Culot, M; Wilmes, A; Schmidt, T; Zurich, M G; Schultz, L; Schmal, O; Gramowski-Voss, A; Weiss, D G; Jennings, P; Bal-Price, A; Testai, E; Dekant, W

    2015-12-25

    There is an increasing need to develop improved systems for predicting the safety of xenobiotics. However, to move beyond hazard identification the available concentration of the test compounds needs to be incorporated. In this study cyclosporine A (CsA) was used as a model compound to assess the kinetic profiles in two rodent brain cell cultures after single and repeated exposures. CsA induced-cyclophilin B (Cyp-B) secretion was also determined as CsA-specific pharmacodynamic endpoint. Since CsA is a potent p-glycoprotein substrate, the ability of this compound to cross the blood-brain barrier (BBB) was also investigated using an in vitro bovine model with repeated exposures up to 14 days. Finally, CsA uptake mechanisms were studied using a parallel artificial membrane assay (PAMPA) in combination with a Caco-2 model. Kinetic results indicate a low intracellular CsA uptake, with no marked bioaccumulation or biotransformation. In addition, only low CsA amounts crossed the BBB. PAMPA and Caco-2 experiments revealed that CsA is mostly trapped to lipophilic compartments and exits the cell apically via active transport. Thus, although CsA is unlikely to enter the brain at cytotoxic concentrations, it may cause alterations in electrical activity and is likely to increase the CNS concentration of other compounds by occupying the BBBs extrusion capacity. Such an integrated testing system, incorporating BBB, brain culture models and kinetics could be applied for assessing neurotoxicity potential of compounds. PMID:25683621

  12. Culture.

    ERIC Educational Resources Information Center

    1997

    Twelve conference papers on cultural aspects of second language instruction include: "Towards True Multiculturalism: Ideas for Teachers" (Brian McVeigh); Comparing Cultures Through Critical Thinking: Development and Interpretations of Meaningful Observations" (Laurel D. Kamada); "Authority and Individualism in Japan and the USA" (Alisa Woodring);…

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

    PubMed Central

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

    1995-01-01

    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, and the DNA was extracted from cells by heat lysis or a sodium iodide-isopropanol or a phenol-chloroform method. DNAs of different sizes were amplified from a region of the MPB70 gene of M. tuberculosis (372 bp) and from a region of the 16S rRNA gene of members of the genus Mycobacterium (1,030 bp), M. intracellulare (850 bp), or M. avium (180 bp) as a multiplex PCR in a single tube. The amplified DNA products were detected by agarose gel electrophoresis and ethidium bromide staining in all 41 (100%) positive cultures after sodium iodide-isopropanol extraction, in 18 (44%) after heat lysis, and in 5 (12%) after phenol-chloroform extraction. Of the 41 positive cultures, 38 were identified as M. avium and 2 were identified as M. intracellulare by both routine methods and multiplex PCR. The remaining mycobacterium was identified as M. intracellulare by routine methods and as M. avium by the multiplex PCR. Another six blood cultures that were negative for the presence of acid-fast bacilli after Ziehl-Neelson staining were also negative by PCR.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7751375

  14. Enrichment of umbilical cord blood mononuclears with hemopoietic precursors in co-culture with mesenchymal stromal cells from human adipose tissue.

    PubMed

    Maslova, E V; Andreeva, E R; Andrianova, I V; Bobyleva, P I; Romanov, Yu A; Kabaeva, N V; Balashova, E E; Ryaskina, S S; Dugina, T N; Buravkova, L B

    2014-02-01

    We demonstrated the possibility of enrichment of umbilical cord blood mononuclear fraction with early non-differentiated precursors under conditions of co-culturing with mesenchymal stromal cells from the human adipose tissue. It was established that umbilical cord blood mononuclear cells adhered to mesenchymal stromal cell feeder and then proliferate and differentiate into hemopoietic cells. In comparison with the initial umbilical cord blood mononuclear fraction, the cell population obtained after 7-day expansion contained 2-fold more CFU and 33.4 ± 9.5 and 24.2 ± 11.2% CD34(+) and CD133(+) cells, respectively, which corresponds to enrichment of precursor cell population by 148 ± 60. The proposed scheme of expansion of hemopoietic cells from umbilical cord blood is economically expedient and can widely used in biology and medicine. PMID:24771453

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

    PubMed

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

    2011-01-01

    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

  16. Microbiological diagnosis of Eggerthella lenta blood culture isolates in a Swedish tertiary hospital: Rapid identification and antimicrobial susceptibility profile.

    PubMed

    Liderot, Karin; Ratcliffe, Paul; Lüthje, Petra; Thidholm, Ellinor; Özenci, Volkan

    2016-04-01

    Eggerthella lenta is a Gram-positive anaerobic bacillus. Improved diagnostics and increased awareness of rare pathogens have revealed its potential to cause serious invasive infections. In this study, 18 clinical E. lenta isolates derived from positive blood cultures were included. Underlying problems of the patients were in the majority of cases related to the gastrointestinal tract. The performance of two MALDI-TOF MS systems, i.e. Bruker and Vitek MS, in identification of E. lenta was analyzed. In addition, the minimal inhibitory concentrations for clinically relevant antimicrobial agents were determined by routine procedures using E-test. 17 of the 18 E. lenta isolates investigated in this study were correctly identified to species level by the Bruker MS system, while the Vitek MS system identified all 18 isolates. Antimicrobial sensitivity towards the tested agents was in general good. However, high resistance rates were observed for penicillin G and piperacillin-tazobactam based on EUCAST breakpoints. PMID:26612006

  17. Reduced susceptibility to vancomycin and biofilm formation in methicillin-resistant Staphylococcus epidermidis isolated from blood cultures

    PubMed Central

    Pinheiro, Luiza; Brito, Carla Ivo; Pereira, Valéria Cataneli; de Oliveira, Adilson; Camargo, Carlos Henrique; da Cunha, Maria de Lourdes Ribeiro de Souza

    2014-01-01

    This study aimed to correlate the presence of ica genes, biofilm formation and antimicrobial resistance in 107 strains of Staphylococcus epidermidis isolated from blood cultures. The isolates were analysed to determine their methicillin resistance, staphylococcal cassette chromosome mec (SCCmec) type, ica genes and biofilm formation and the vancomycin minimum inhibitory concentration (MIC) was measured for isolates and subpopulations growing on vancomycin screen agar. The mecA gene was detected in 81.3% of the S. epidermidis isolated and 48.2% carried SCCmec type III. The complete icaADBC operon was observed in 38.3% of the isolates; of these, 58.5% produced a biofilm. Furthermore, 47.7% of the isolates grew on vancomycin screen agar, with an increase in the MIC in 75.9% of the isolates. Determination of the MIC of subpopulations revealed that 64.7% had an MIC ≥ 4 μg mL-1, including 15.7% with an MIC of 8 μg mL-1 and 2% with an MIC of 16 μg mL-1. The presence of the icaADBC operon, biofilm production and reduced susceptibility to vancomycin were associated with methicillin resistance. This study reveals a high level of methicillin resistance, biofilm formation and reduced susceptibility to vancomycin in subpopulations of S. epidermidis. These findings may explain the selection of multidrug-resistant isolates in hospital settings and the consequent failure of antimicrobial treatment. PMID:25410990

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

    PubMed

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

    2015-12-01

    The aim of this study is to compare conventional methods, CHROMagar Candida, VITEK2 YST card and VITEK®MS system for the identification of Candida strains isolated from blood cultures. Fifty-four strains were identified according to conventional methods, CHROMagar Candida, VITEK2 YST card and VITEK®MS. Sequencing was used as the reference method. The 54 strains included 32 Candida parapsilosis, 19 Candida albicans, 1 Candida glabrata and 2 Candida tropicalis according to the reference method. One C. albicans and one C. glabrata isolate were misidentified as C. parapsilosis by CHROMagar Candida. Two C. parapsilosis and three C. albicans isolates were misidentified by VITEK2 YST card. CHROMagar Candida, VITEK2 YST card and VITEK®MS identified correctly 96.2%, 90.7% and 100% of all strains, respectively. We found that the CHROMagar Candida, VITEK2 YST card and VITEK®MS system are easy, rapid and accurate alternative methods for the identification of yeast species in the clinical microbiology laboratory. PMID:26700081

  19. Aerobic microorganism for the degradation of chlorinated aliphatic hydrocarbons

    DOEpatents

    Fliermans, Carl B.

    1989-01-01

    A chlorinated aliphatic hydrocarbon-degrading microorganism, having American Type Culture Collection accession numbers ATCC 53570 and 53571, in a biologically pure culture aseptically collected from a deep subsurface habitat and enhanced, mineralizes trichloroethylene and tetrachloroethylene to HCl, H.sub.2 O and Co.sub.2 under aerobic conditions stimulated by methane, acetate, methanol, tryptone-yeast extract, propane and propane-methane.

  20. Rapid detection of bacterial growth in blood samples by a continuous-monitoring electrical impedance apparatus.

    PubMed Central

    Specter, S; Throm, R; Strauss, R; Friedman, H

    1977-01-01

    A growth detection method utilizing an automated apparatus capable of rapidly detecting bacterial growth by measuring changes of electrical impedance in bacteriological medium was utilized with "mock" blood cultures containing various gram-negative and gram-positive bacteria. Measurement of changes of electrical impedance was found to ba as accurate and comparable for time of growth detection as the radiometric method for detection of the same bacteria using mock blood cultures. In a limited clinical trial the use of the electrical impedance apparatus detected in 1 positive specimen from 40 clinical blood specimens as rapidly as by radiometric measurement. Both methods were considerably faster for detecting bacterial growth as compared with conventional culture methods. The selected species of gram-positive and -negative organisms tested were all detected by the electrical impedance method, including aerobes and anerobes. However, addition of 5% CO2 to the incubation atmosphere enhanced detection of gram-positive organisms. PMID:336642

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

    PubMed Central

    Morgenthaler, Nils G.; Kostrzewa, Markus

    2015-01-01

    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

  2. Expression of human immunodeficiency virus (HIV) in naturally infected peripheral blood mononuclear cells: comparison of a standard co-culture technique with a newly developed microculture method.

    PubMed

    Eberlein, B; Baur, A; Neundorfer, M; Jahn, G

    1991-05-01

    Peripheral blood mononuclear cells (PBMCs) from 29 patients infected with human immunodeficiency virus (HIV) were cultured by two different methods. One was the standard co-culture technique, the other a newly developed microculture method. In this assay 10(6) PBMCs were cultivated in 250 microliters medium, no activating agents or allogeneic cells were present. P24 antigen production measured by this method was found in 7 out of 11 PBMC cultures of patients in the Walter Reed (WR) stage 1 or 2, whereas only 4 samples were positive by the co-culture procedure. Cultures from patients in the later stages of the disease (WR 5/6) showed a higher p24 production by the co-culture method than by the microculture assay. It is assumed that rapidly growing HIV strains can be better assessed by the co-culture method which may select for these strains. P24 expression can be more easily obtained by the microculture technique even in cases where slowly replicating strains may be present. In conclusion, results from the microculture procedure described may be a useful supplementation to findings observed by the co-culture method. PMID:1909827

  3. Evaluation of fluorescence in situ hybridisation (FISH) for the detection of fungi directly from blood cultures and cerebrospinal fluid from patients with suspected invasive mycoses.

    PubMed

    Da Silva, Roberto Moreira; Da Silva Neto, João Ricardo; Santos, Carla Silvana; Frickmann, Hagen; Poppert, Sven; Cruz, Kátia Santana; Koshikene, Daniela; De Souza, João Vicente Braga

    2015-01-01

    The aim of this study was to evaluate the diagnostic performance of in-house FISH (fluorescence in situ hybridisation) procedures for the direct identification of invasive fungal infections in blood cultures and cerebrospinal fluid (CSF) samples and to compare these FISH results with those obtained using traditional microbiological techniques and PCR targeting of the ITS1 region of the rRNA gene. In total, 112 CSF samples and 30 positive blood cultures were investigated by microscopic examination, culture, PCR-RFLP and FISH. The sensitivity of FISH for fungal infections in CSF proved to be slightly better than that of conventional microscopy (India ink) under the experimental conditions, detecting 48 (instead of 46) infections in 112 samples. The discriminatory powers of traditional microbiology, PCR-RFLP and FISH for fungal bloodstream infections were equivalent, with the detection of 14 fungal infections in 30 samples. However, the mean times to diagnosis after the detection of microbial growth by automated blood culture systems were 5 hours, 20 hours and 6 days for FISH, PCR-RFLP and traditional microbiology, respectively. The results demonstrate that FISH is a valuable tool for the identification of invasive mycoses that can be implemented in the diagnostic routine of hospital laboratories. PMID:25637361

  4. Comparative analysis of Gram's stain, PNA-FISH and Sepsityper with MALDI-TOF MS for the identification of yeast direct from positive blood cultures.

    PubMed

    Gorton, Rebecca L; Ramnarain, P; Barker, K; Stone, N; Rattenbury, S; McHugh, T D; Kibbler, C C

    2014-10-01

    Fungaemia diagnosis could be improved by reducing the time to identification of yeast from blood cultures. This study aimed to evaluate three rapid methods for the identification of yeast direct from blood cultures; Gram's stain analysis, the AdvanDX Peptide Nucleic Acid in Situ Hybridisation Yeast Traffic Light system (PNA-FISH YTL) and Bruker Sepsityper alongside matrix-assisted laser desorption ionisation time of flight mass spectrometry (MALDI-TOF MS). Fifty blood cultures spiked with a known single yeast strain were analysed by blinded operators experienced in each method. Identifications were compared with MALDI-TOF MS CHROMagar Candida culture and ITS rRNA sequence-based identifications. On first attempt, success rates of 96% (48/50) and 76% (36/50) were achieved using PNA-FISH YTL and Gram's stain respectively. MALDI-TOF MS demonstrated a success rate of 56% (28/50) when applying manufacturer's species log score thresholds and 76% (38/50) using in-house parameters, including lowering the species log score threshold to >1.5. In conclusion, PNA-FISH YTL demonstrated a high success rate successfully identifying yeast commonly encountered in fungaemia. Sepsityper(™) with MALDI-TOF MS was accurate but increased sensitivity is required. Due to the misidentification of commonly encountered yeast Gram's stain analysis demonstrated limited utility in this setting. PMID:24862948

  5. Characterization and aerobic biodegradation of selected monoterpenes

    SciTech Connect

    Misra, G.; Pavlostathis, S.G.; Li, J.; Purdue, E.M.

    1996-12-31

    Monoterpenes are biogenic chemicals and occur in abundance in nature. Large-scale industrial use of these chemicals has recently been initiated in an attempt to replace halogenated solvents and chlorofluorocarbons which have been implicated in the stratospheric depletion of ozone. This study examined four hydrocarbon monoterpenes (d-limonene, {alpha}-pinene, {gamma}-terpinene, and terpinolene) and four alcohols (arbanol, linalool, plinol, and {alpha}-terpineol). Water solubility, vapor pressure, and octanol/water partition coefficients were estimated. Aerobic biodegradability tests were conducted in batch reactors by utilizing forest soil extract and enriched cultures as inoculum. The hydrophobic nature and high volatility of the hydrocarbons restricted the investigation to relatively low aqueous concentrations. Each monoterpene was analyzed with a gas chromatograph equipped with a flame ionization detector after extraction from the aqueous phase with isooctane. Terpene mineralization was tested by monitoring liquid-phase carbon, CO{sub 2} production and biomass growth. All four hydrocarbons and two alcohols readily degraded under aerobic conditions. Plinol resisted degradation in assays using inocula from diverse sources, while arbanol degraded very slowly. The intrinsic biokinetics coefficients for the degradation of d-limonene and {alpha}-terpineol were estimated by using cultures enriched with the respective monoterpenes. Monoterpene biodegradation followed Monod kinetics.

  6. Evaluation of the Punch-it™ NA-Sample kit for detecting microbial DNA in blood culture bottles using PCR-reverse blot hybridization assay.

    PubMed

    Kim, Jungho; Wang, Hye-Young; Kim, Seoyong; Park, Soon Deok; Yu, Kwangmin; Kim, Hyo Youl; Uh, Young; Lee, Hyeyoung

    2016-09-01

    DNA extraction efficiency affects the success of PCR-based method applications. The Punch-it™ NA-Sample kit for extracting DNA by using paper chromatography is technically easy to use and requires just two reagents and only 10min to complete. The Punch-it™ NA-Sample kit could be offered as a rapid, accurate, and convenient method for extracting bacterial and fungal DNA from blood culture bottles. We compared the efficiencies of the commercial kit (Punch-it™ NA-Sample kit) and an in-house conventional boiling method with Chelex-100 resin for DNA extraction from blood culture bottles. The efficiency of the two DNA extraction methods was assessed by PCR-reverse blot hybridization assay (PCR-REBA, REBA Sepsis-ID) for detecting Gram positive (GP) bacteria, Gram negative (GN) bacteria, and Candida species with 196 positive and 200 negative blood culture bottles. The detection limits of the two DNA extraction methods were 10(3)CFU/mL for GP bacteria, 10(3)CFU/mL for GN bacteria, and 10(4)CFU/mL for Candida. The sensitivity and specificity of the Punch-it™ NA-Sample kit by REBA Sepsis-ID were 95.4% (187/196) and 100% (200/200), respectively. The overall agreement of the two DNA extraction methods was 98.9% (392/396). Three of four samples showing discrepant results between the two extraction methods were more accurately matched up with the Punch-it™ NA-Sample kit based on conventional culture methods. The results indicated that the Punch-it™ NA-Sample kit extracted bacterial and fungal DNA in blood culture bottles and allowed extracted DNA to be used in molecular assay. PMID:27263831

  7. Field tests for evaluating the aerobic work capacity of firefighters.

    PubMed

    Lindberg, Ann-Sofie; Oksa, Juha; Gavhed, Désirée; Malm, Christer

    2013-01-01

    Working as a firefighter is physically strenuous, and a high level of physical fitness increases a firefighter's ability to cope with the physical stress of their profession. Direct measurements of aerobic capacity, however, are often complicated, time consuming, and expensive. The first aim of the present study was to evaluate the correlations between direct (laboratory) and indirect (field) aerobic capacity tests with common and physically demanding firefighting tasks. The second aim was to give recommendations as to which field tests may be the most useful for evaluating firefighters' aerobic work capacity. A total of 38 subjects (26 men and 12 women) were included. Two aerobic capacity tests, six field tests, and seven firefighting tasks were performed. Lactate threshold and onset of blood lactate accumulation were found to be correlated to the performance of one work task (r(s) = -0.65 and -0.63, p<0.01, respectively). Absolute (mL · min(-1)) and relative (mL · kg(-1) · min(-1)) maximal aerobic capacity was correlated to all but one of the work tasks (r(s) = -0.79 to 0.55 and -0.74 to 0.47, p<0.01, respectively). Aerobic capacity is important for firefighters' work performance, and we have concluded that the time to row 500 m, the time to run 3000 m relative to body weight (s · kg(-1)), and the percent of maximal heart rate achieved during treadmill walking are the most valid field tests for evaluating a firefighter's aerobic work capacity. PMID:23844153

  8. Classification of positive blood cultures: computer algorithms versus physicians' assessment - development of tools for surveillance of bloodstream infection prognosis using population-based laboratory databases

    PubMed Central

    2012-01-01

    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- or hospital-onset; d) community-onset bloodstream infection as healthcare-associated or not. We applied the computer algorithms to data from laboratory databases and the Danish National Patient Registry to classify the same groups and compared these with the physicians’ assessments as reference episodes. For each classification, we tabulated episodes derived by the physicians’ assessment and the computer algorithm and compared 30-day mortality between concordant and discrepant groups with adjustment for age, gender, and comorbidity. Results Physicians derived 9,482 reference episodes from 21,705 positive blood cultures. The agreement between computer algorithms and physicians’ assessments was high for contamination vs. bloodstream infection (8,966/9,482 reference episodes [96.6%], Kappa = 0.83) and mono- vs. polymicrobial bloodstream infection (6,932/7,288 reference episodes [95.2%], Kappa = 0.76), but lower for community- vs. hospital-onset bloodstream infection (6,056/7,288 reference episodes [83.1%], Kappa = 0.57) and healthcare-association (3

  9. Efficiency of transgenic T cell generation from gene-marked cultured human CD34+ cord blood cells is determined by their maturity and the cytokines present in the culture medium.

    PubMed

    Verhasselt, B; Naessens, E; De Smedt, M; Plum, J

    2000-05-01

    Success of gene therapy for diseases affecting the T cell lineage depends on the thymic repopulation by genetically engineered hematopoietic progenitor cells (HPC). Although it has been shown that retrovirally transduced HPC can repopulate the thymus, little information is available on the effect of the culture protocol. Moreover, for expansion of the number of HPC, cytokine supplemented culture is needed. Here, we transduced purified human umbilical cord blood (CB) CD34+ cells in cultures supplemented with various combinations of the cytokines thrombopoietin (TPO), stem cell factor (SCF), flt3/flk-2 ligand (FL), interleukin-3 (IL-3) and IL-6, and investigated thymus-repopulating ability of gene-marked HPC in vitro. Irrespective of the cytokine cocktail used, transduced CD34+CD38- CB cells, expressing the marker green fluorescent protein (GFP) encoded by the MFG-GFP retrovirus, have both superior proliferative and thymus-repopulating potential compared with transduced CD34+CD38+ CB cells. Effectively transduced GFP+CD34+CD38- HPC, cultured for 3 or 17 days, more readily generated T cells than GFP- HPC from the same culture. The reverse was true in the case of CD34+CD38+ HPC cultures. Finally, our results indicate that the number of GFP+ T cell progenitors actually increased during culture of CD34+CD38- HPC, in a magnitude that is determined by the cytokine cocktail used during culture. PMID:10845720

  10. Blood Culture (For Parents)

    MedlinePlus

    ... Diseases & Conditions Pregnancy & Baby Nutrition & Fitness Emotions & Behavior School & Family Life First Aid & Safety Doctors & Hospitals Q& ... What to Say Vaccines: Which Ones & When? Smart School Lunches Emmy-Nominated Video "Cerebral Palsy: Shannon's Story" ...

  11. [Characterization and determination of antibiotic resistance profiles of a single clone Acinetobacter baumannii strains isolated from blood cultures].

    PubMed

    Karagöz, Alper; Baran, Irmak; Aksu, Neriman; Acar, Sümeyra; Durmaz, Rıza

    2014-10-01

    Acinetobacter baumannii which is a significant cause of nosocomial infections, increases the rate of morbidity and mortality in health care settings especially in intensive care units (ICUs). The aim of this study was to determine the antibiotic resistance profiles of A.baumannii strains isolated from blood cultures of inpatients from different ICUs, wards and hospital environment and evaluate their clonal relationships and epidemiologic features. A total of 54 A.baumannii strains (47 from the blood cultures and 7 from the hospital environment), identified between 01 January 2012-28 December 2012 at the Clinical Microbiology Laboratory of Ankara Numune Training and Research Hospital, Turkey, were included in the study. Identification of A.baumannii isolates and their antimicrobial [sulbactam-ampicillin (SAM), piperacillin (PIP), piperacillin-tazobactam (TZP), ceftazidime (CFZ), cefoperazone-sulbactam (SCF), cefepime (CEF), imipenem (IMP), meropenem (MER), amikacin (AMK), gentamicin (GEN), netilmicin (NT), ciprofloxacin (CIP), levofloxacin (LVF), tetracycline (TET), tigecycline (TG), colistin (COL), trimethoprim-sulfamethoxazole (SXT)] susceptibility testing were performed by Vitek 2 (bioMérieux, France) system. The clonal relationship between the A.baumannii isolates was analysed by pulsed-field gel electrophoresis (PFGE). In our study colistin, tigecycline and netilmicin were found to be the most effective agents against A.baumannii isolates. All of the clinical isolates (n= 47) were found susceptible to COL, however all were resistant to SAM, PIP, TZP, CEF, IPM, CFZ, MER and CIP. While 1.85%, 14.8%, 14.8%, 16.6%, 59.2% and 22.2% of the isolates were susceptible to SCF, AMK, NT, GEN, TG and SXT, respectively; 1.85%, 1.85%, 9.2%, 16.6%, 38.8% and 27.7% of the isolates were intermediate to SCF, TET, AMK, NT, LVF and TG, respectively. Similarly, all of the environmental A.baumannii isolates (n= 7) were resistant to SAM, PIP, TZP, CFZ, CEF, IPM, MER and CIP, and all

  12. Aerobic and anaerobic microbiology of infections after trauma in children.

    PubMed Central

    Brook, I

    1998-01-01

    OBJECTIVE: To review the recovery of aerobic and anaerobic bacteria from infections after trauma in children over a 20 year period. METHODS: Only specimens that were studied for both aerobic and anaerobic bacteria were included in the analysis. They were collected from seven separate centres in which the microbiology laboratories only accepted specimens that were properly collected without contamination and were submitted in appropriate transport media. Anaerobes and aerobic bacteria were cultured and identified using standard techniques. Clinical records were reviewed to identify post-trauma patients. RESULTS: From 1974 to 1994, 175 specimens obtained from 166 children with trauma showed bacterial growth. The trauma included blunt trauma (71), lacerations (48), bites (42), and open fractures (5). Anaerobic bacteria only were isolated in 38 specimens (22%), aerobic bacteria only in 51 (29%), and mixed aerobic-anaerobic flora in 86 (49%); 363 anaerobic (2.1/specimen) and 158 aerobic or facultative isolates (0.9/specimen) were recovered. The predominant anaerobic bacteria included Peptostreptococcus spp (115 isolates), Prevotella spp (68), Fusobacterium spp (52), B fragilis group (42), and Clostridium spp (21). The predominant aerobic bacteria included Staph aureus (51), E coli (13), Ps aeruginosa (12), Str pyogenes (11) and Klebsiella pneumoniae (9). Principal infections were: abscesses (52), bacteraemia (3), pulmonary infections (30, including aspiration pneumonia, tracheostomy associated pneumonia, empyema, and ventilator associated pneumonia), wounds (36, including cellulitis, post-traumatic wounds, decubitus ulcers, myositis, gastrostomy and tracheostomy site wounds, and fasciitis), bites (42, including 23 animal and 19 human), peritonitis (4), osteomyelitis (5), and sinusitis (3). Staph aureus and Str pyogenes were isolated at all sites. However, organisms of the oropharyngeal flora predominated in infections that originated from head and neck wounds and

  13. Staphylococcus pseudolugdunensis sp. nov., a pyrrolidonyl arylamidase/ornithine decarboxylase-positive bacterium isolated from blood cultures.

    PubMed

    Tang, Yi-Wei; Han, Jian; McCormac, Melinda A; Li, Haijing; Stratton, Charles W

    2008-04-01

    Twelve coagulase-negative staphylococcal isolates recovered from blood cultures with positive pyrrolidonyl arylamidase and ornithine decarboxylase reactions were identified as Staphylococcus lugdunensis by the clinical microbiology laboratory. However, none of these 12 isolates were recognized by a S. lugdunensis translation elongation factor Tu (tuf) gene-specific probe. Under the API STAPH V4.0 identification system (bioMérieux, Durham, NC), 8 of these 12 isolates could not be identified with low discrimination scores, and 4 were identified as Kocuria varians/rosea with identification probabilities that ranged from 95.5% to 99.6%. All 12 isolates possessed identical partial 16S rRNA gene sequences, and the full 16S rRNA gene sequences of the prototype strain B006 were closely related to a tentatively assigned "Staphylococcus pettenkoferi". All 12 isolates had identical partial tuf gene sequences corresponding to 666 to 858 nucleotide positions, and the 1188-base pair full tuf sequences of B006 strain were mostly related to 2 Staphylococcus epidermidis isolates with a 93.02% similarity. Two isolates, which were recovered from the same patient over a 16-day interval, were considered to be a pathogen causing an intravenous line-associated infection; the remaining 10 isolates were considered to be skin contaminants. Biochemical tests currently used in the clinical microbiology laboratory to identify S. lugdunensis appear to lack specificity in identifying these isolates. On the basis of the close biochemical reactions with S. lugdunensis and phylogenetic evidence, these isolates are proposed the designation Staphylococcus pseudolugdunensis sp. nov. PMID:18248933

  14. Proinflammatory effect of trivalent arsenical species in a co-culture of Caco-2 cells and peripheral blood mononuclear cells.

    PubMed

    Calatayud, Marta; Gimeno-Alcañiz, José V; Devesa, Vicenta; Vélez, Dinoraz

    2015-04-01

    Chronic exposure to inorganic arsenic (As) is associated with type 2 diabetes, cardiovascular diseases and cancer. Ingested inorganic As is transformed within the gastrointestinal tract and can give rise to more toxic species such as monomethylarsonous acid [MMA(III)] and dimethylarsinous acid [DMA(III)]. Thus, the intestinal epithelium comes into contact with toxic arsenical species, and the effects of such exposure upon epithelial function are not clear. The present study has evaluated the effect of 1 µM arsenite [As(III)], 0.1 µM MMA(III) and 1 µM DMA(III) upon the release of cytokines [interleukin-6 (IL6), IL8, tumor necrosis factor alpha (TNFα)], using a compartmentalized co-culture model with differentiated Caco-2 cells in the apical compartment and peripheral blood mononuclear cells in the basolateral compartment. In addition, the combined effect of arsenical species and lipopolysaccharide (LPS), both added into the apical compartment, has been analyzed. The results indicate that exposure to the arsenical forms induces a proinflammatory response. An increase in cytokine secretion into the basolateral compartment was observed, particularly as regards TNFα (up to 1,600 %). The cytokine levels on the apical side also increased, though to a lesser extent. As/LPS co-exposure significantly affected the proinflammatory response as compared to treatment with As alone. Treatment with DMA(III) and As/LPS co-exposure increased the permeability of the intestinal monolayer. In addition, As/LPS treatments enhanced As(III) and MMA(III) transport through the intestinal monolayer. PMID:24862236

  15. Blood Culture Proven Early Onset Sepsis and Late Onset Sepsis in Very-Low-Birth-Weight Infants in Korea

    PubMed Central

    Lee, Soon Min; Chang, Meayoung

    2015-01-01

    Neonatal sepsis remains one of the most important causes of death and co-morbidity in very-low-birth-weight (VLBW) infants. The aim of this study was to determine the current incidences of early-onset sepsis (EOS) and late-onset sepsis (LOS), the distribution of pathogens, and the impact of infection on co-morbidities in VLBW infants. We analyzed the data including sepsis episode from 2,386 VLBW infants enrolled in Korean Neonatal Network from January 2013 to June 2014. We defined EOS as a positive blood culture occurring between birth and 7 days of life and LOS after 7 days of life. Sepsis was found in 21.1% of VLBW infants. The risk of sepsis was inversely related to birth weight and gestational age. EOS was found in only 3.6% of VLBW infants, however the mortality rate was as high as 34.1%. EOS was associated with the increased odds for bronchopulmonary dysplasia and intraventricular hemorrhage. The vast majority of EOS was caused by Gram-positive organisms, particularly coagulase-negative staphylococci (30.6%). LOS developed in 19.4% of VLBW infants with a 16.1% mortality rate. Pathogens in LOS were dominated by coagulase-negative staphylococci (38.3%). Twenty-five percent and fifty percent of first LOS episode occurred after 12 days and 20 days from birth, respectively. Younger and smaller VLBW infants showed the earlier occurrence day for the 25% of first LOS episode. This study provides a recent nationwide epidemiology of sepsis in VLBW infants in Korea. Based on this study, successful strategies to reduce infections would improve survival and reduce morbidity. PMID:26566360

  16. Pericytes support neutrophil transmigration via interleukin-8 across a porcine co-culture model of the blood-brain barrier.

    PubMed

    Pieper, Christian; Pieloch, Paulina; Galla, Hans-Joachim

    2013-08-01

    Transmigration of neutrophils across the blood-brain barrier (BBB) to an inflamed brain tissue is an important process during neuronal inflammation. The process of neutrophil activation as well as their way of rolling along the endothelium and their transmigration is quite well understood. Nevertheless, relatively little is known about the fate of neutrophils after they have transmigrated through the endothelium. The role of the other cells of the neurovascular unit in this process is also poorly understood. Here we studied the potential of pericytes to chemo-attract neutrophils under inflammatory conditions. Quantitative real time PCR, western blot analysis, and a chemotaxis assay showed that pericytes are able to chemo-attract neutrophils by interleukin-8 (IL-8) after stimulation with lipopolysaccharides (LPS), tumor necrosis factor-alpha (TNF-α), or interleukin-1beta (IL-1β). Then, a co-culture model consisting of primary porcine brain capillary endothelial cells (PBCECs) and primary porcine brain capillary pericytes (PBCPs) was used to analyze neutrophil transmigration across the BBB. As a model for inflammation, LPS was used and the effects of the cytokines TNF-α, IL-1β, and interferon-gamma (IFN-γ) were analyzed. In general, all stimulants apart from IFN-γ were able to increase transendothelial neutrophil migration. This effect was significantly reduced by a specific inhibitor of matrix metalloproteinases (MMPs)-2 and -9. MMP-2/-9 secretion is expected to decrease adhesion to pericytes and thus support the transmigration of neutrophils. Additionally, in an adhesion experiment, we showed that MMP-2/-9 inhibition significantly enhances the adhesion of neutrophils to pericytes. PMID:23769734

  17. Blood culture confirmed bacterial sepsis in neonates in a North Indian tertiary care center: changes over the last decade.

    PubMed

    Sundaram, Venkataseshan; Kumar, Praveen; Dutta, Sourabh; Mukhopadhyay, Kanya; Ray, Pallab; Gautam, Vikas; Narang, Anil

    2009-01-01

    The spectrum of organisms causing sepsis is different in developing countries. Data on the recent trends of organisms causing sepsis are limited. This study was conducted in a tertiary care neonatal unit in Northern India. All inborn babies with blood-culture-positive sepsis from 1995 to 2006 were divided into two epochs, viz. 1995 to 1998 (epoch I) and 2001 to 2006 (epoch II). Organisms were grouped into early (<72 h) and late onset (> or =72 h) sepsis groups. The overall incidence of sepsis, the incidence of sepsis stratified by weight groups, the organism profile on different days of life, sepsis-related mortality and pathogen-specific case fatality rate were calculated and compared between the two epochs. Out of 34,362 live births during the study period, organisms were isolated in 1,491 neonates. Out of these, 89% had bacterial sepsis. The incidence of neonatal bacterial sepsis increased from epoch I to epoch II (35.8/1,000 versus 40.1/1,000 live births, P<0.05). The incidence of early onset sepsis (EOS) did not change between the epochs, but the incidence of late onset sepsis (LOS) increased from 12 to 16.5 per 1,000 live births (P<0.001). The incidence of bacterial sepsis decreased significantly in the 1,000- to 1,999-g birth weight groups. Klebsiella pneumoniae and Enterobacter aerogenes decreased, whereas Staphylococcus aureus increased in incidence during epoch II. Non-fermenting Gram-negative bacilli emerged as a newly identified pathogen during epoch II. Sepsis-associated mortality decreased from 42 to 20%. The incidence of bacterial sepsis has decreased significantly in 1,000- to 1,999-g infants, with a significant reduction in sepsis-related mortality. New organisms have emerged in recent years. The organism profile in recent years has changed, with a significant overlap of organisms causing EOS and LOS. PMID:19168958

  18. Aerobic and anaerobic exercise training in obese adults

    PubMed Central

    Al Saif, Amer; Alsenany, Samira

    2015-01-01

    [Purpose] Obesity is a global health problem and is associated with a multitude of complications. This study was designed to determine changes in cardiopulmonary functions after aerobic and anaerobic exercise training in obese subjects. [Subjects and Methods] Forty obese subjects, whose ages ranged between 18 and 25 years, were divided into 2 equal groups: group A received aerobic exercise training in addition to dietary measures, and group B received anaerobic exercise training for 3 months in addition to dietary measures. Measurements of systolic blood pressure, diastolic blood pressure, heart rate, maximum voluntary ventilation, maximal oxygen consumption, and body mass index were obtained for both groups before and after the exercise program. [Results] The mean body mass index, systolic blood pressure, diastolic blood pressure, heart rate, and maximal oxygen consumption decreased significantly, whereas the mean maximum voluntary ventilation increased significantly after treatment in group A. The mean maximum voluntary ventilation also increased significantly after treatment in group B. There were significant differences between the mean levels of the investigated parameters in groups A and B after treatment. [Conclusion] Aerobic exercise reduces weight and improves cardiopulmonary fitness in obese subjects better than anaerobic exercise. PMID:26180300

  19. Design of a Tablet Computer App for Facilitation of a Molecular Blood Culture Test in Clinical Microbiology and Preliminary Usability Evaluation

    PubMed Central

    Pape-Haugaard, Louise; Meltzer, Michelle C; Fuchs, Martin; Schønheyder, Henrik C; Hejlesen, Ole

    2016-01-01

    Background User mobility is an important aspect of the development of clinical information systems for health care professionals. Mobile phones and tablet computers have obtained widespread use by health care professionals, offering an opportunity for supporting the access to patient information through specialized applications (apps) while supporting the mobility of the users. The use of apps for mobile phones and tablet computers may support workflow of complex tasks, for example, molecular-based diagnostic tests in clinical microbiology. Multiplex Blood Culture Test (MuxBCT) is a molecular-based diagnostic test used for rapid identification of pathogens in positive blood cultures. To facilitate the workflow of the MuxBCT, a specialized tablet computer app was developed as an accessory to the diagnostic test. The app aims to reduce the complexity of the test by step-by-step guidance of microscopy and to assist users in reaching an exact bacterial or fungal diagnosis based on blood specimen observations and controls. Additionally, the app allows for entry of test results, and communication thereof to the laboratory information system (LIS). Objective The objective of the study was to describe the design considerations of the MuxBCT app and the results of a preliminary usability evaluation. Methods The MuxBCT tablet app was developed and set up for use in a clinical microbiology laboratory. A near-live simulation study was conducted in the clinical microbiology laboratory to evaluate the usability of the MuxBCT app. The study was designed to achieve a high degree of realism as participants carried out a scenario representing the context of use for the MuxBCT app. As the MuxBCT was under development, the scenario involved the use of molecular blood culture tests similar to the MuxBCT for identification of microorganisms from positive blood culture samples. The study participants were observed, and their interactions with the app were recorded. After the study, the

  20. Aerobic Denitrifying Bacteria That Produce Low Levels of Nitrous Oxide

    PubMed Central

    Takaya, Naoki; Catalan-Sakairi, Maria Antonina B.; Sakaguchi, Yasushi; Kato, Isao; Zhou, Zhemin; Shoun, Hirofumi

    2003-01-01

    Most denitrifiers produce nitrous oxide (N2O) instead of dinitrogen (N2) under aerobic conditions. We isolated and characterized novel aerobic denitrifiers that produce low levels of N2O under aerobic conditions. We monitored the denitrification activities of two of the isolates, strains TR2 and K50, in batch and continuous cultures. Both strains reduced nitrate (NO3−) to N2 at rates of 0.9 and 0.03 μmol min−1 unit of optical density at 540 nm−1 at dissolved oxygen (O2) (DO) concentrations of 39 and 38 μmol liter−1, respectively. At the same DO level, the typical denitrifier Pseudomonas stutzeri and the previously described aerobic denitrifier Paracoccus denitrificans did not produce N2 but evolved more than 10-fold more N2O than strains TR2 and K50 evolved. The isolates denitrified NO3− with concomitant consumption of O2. These results indicated that strains TR2 and K50 are aerobic denitrifiers. These two isolates were taxonomically placed in the β subclass of the class Proteobacteria and were identified as P. stutzeri TR2 and Pseudomonas sp. strain K50. These strains should be useful for future investigations of the mechanisms of denitrifying bacteria that regulate N2O emission, the single-stage process for nitrogen removal, and microbial N2O emission into the ecosystem. PMID:12788710

  1. Impact of a Rapid Microarray-Based Assay for Identification of Positive Blood Cultures for Treatment Optimization for Patients with Streptococcal and Enterococcal Bacteremia

    PubMed Central

    Roshdy, Danya G.; Tran, Anthony; LeCroy, Nicholas; Zeng, Donglin; Ou, Fang-shu; Daniels, Lindsay M.; Weber, David J.; Alby, Kevin

    2015-01-01

    Implementation of the Verigene Gram-positive blood culture test led to reductions in time to acceptable antibiotic overall (1.9 versus 13.2 h, respectively; P = 0.04) and time to appropriate antibiotic for patients with vancomycin-resistant Enterococcus (4.2 versus 43.7 h; P = 0.006) and viridans group Streptococcus (0.2 versus 7.1 h; P = 0.02). PMID:25673785

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

    PubMed Central

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

    2012-01-01

    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 bacilli identified by Gram staining, we isolated bacterial DNA using spin columns (BC-C) and rapid water lysis (BC-W). Twenty ESBL/KPC-positive and 20 ESBL/KPC-negative blood culture samples, as well as 20 non-lactose-fermenting organisms, were tested. The 20 isolates that were ESBL positive by phenotypic testing were also evaluated on solid medium (SM), and the DNA was extracted by use of a spin column (SM-C). The resulting 140 DNA extractions were assessed for DNA quantity and quality using 260/280-nm absorbance ratios, and DNA microarray analysis was performed in a blinded fashion. Microarray and phenotypic results were concordant for 98.3% of BC-W, 90% of BC-C, and 95% of SM-C samples. Compared to phenotypic testing, the sensitivity and specificity for BC-C samples were 88.9% and 100%, respectively, and for BC-W samples, the sensitivity and specificity were 94.4% and 100%, respectively. BC-W samples yielded the highest concordance with phenotypic results. Nucleic acid microarrays offer promise in the identification of blaESBL and blaKPC genes directly from blood cultures, thereby reducing the time to identification of these important pathogens. PMID:22718942

  3. Early detection of extended-spectrum β-lactamase from blood culture positive for an Enterobacteriaceae using βLACTA test

    PubMed Central

    Prod'hom, Guy; Durussel, Christian; Blanc, Dominique; Croxatto, Antony; Greub, Gilbert

    2015-01-01

    Bacterial pellets from Enterobacteriaceae positive blood cultures prepared using ammonium chloride were tested for rapid detection of β-lactamase using the commercial βLACTA test and read after 30 minutes. During 7 months, 137 bacterial pellets were tested prospectively. βLACTA test exhibited a sensitivity of 75% and a specificity of 100% for the detection of third-generation cephalosporin resistance. False negative tests were mainly observed with hyperproduced chromosomal or plasmid-borne AmpC. PMID:26380714

  4. Influence of aerobic and anoxic microenvironments on polyhydroxyalkanoates (PHA) production from food waste and acidogenic effluents using aerobic consortia.

    PubMed

    Reddy, M Venkateswar; Mohan, S Venkata

    2012-01-01

    The functional role of aerobic and anoxic microenvironments on polyhydroxyalkanoates (PHA) production using food waste (UFW) and effluents from acidogenic biohydrogen production process (FFW) were studied employing aerobic mixed culture as biocatalyst. Anoxic microenvironment documented higher PHA production, while aerobic microenvironment showed higher substrate degradation. FFW showed higher PHA accumulation (39.6%) than UFW (35.6%) due to ready availability of precursors (fatty acids). Higher fraction of poly-3-hydroxy butyrate (PHB) was observed compared to poly-3-hydroxy valerate (PHV) in the accumulated PHA in the form of co-polymer [P3(HB-co-HV)]. Dehydrogenase, phosphatase and protease enzymatic activities were monitored during process operation. Integration with fermentative biohydrogen production yielded additional substrate degradation under both aerobic (78%) and anoxic (72%) microenvironments apart from PHA production. Microbial community analysis documented the presence of aerobic and facultative organisms capable of producing PHA. Integration strategy showed feasibility of producing hydrogen along with PHA by consuming fatty acids generated during acidogenic process in association with increased treatment efficiency. PMID:22055090

  5. Sequence-Based Identification of Aerobic Actinomycetes

    PubMed Central

    Patel, Jean Baldus; Wallace, Richard J.; Brown-Elliott, Barbara A.; Taylor, Tony; Imperatrice, Carol; Leonard, Deborah G. B.; Wilson, Rebecca W.; Mann, Linda; Jost, Kenneth C.; Nachamkin, Irving

    2004-01-01

    We investigated the utility of 500-bp 16S rRNA gene sequencing for identifying clinically significant species of aerobic actinomycetes. A total of 28 reference strains and 71 clinical isolates that included members of the genera Streptomyces, Gordonia, and Tsukamurella and 10 taxa of Nocardia were studied. Methods of nonsequencing analyses included growth and biochemical analysis, PCR-restriction enzyme analysis of the 439-bp Telenti fragment of the 65 hsp gene, susceptibility testing, and, for selected isolates, high-performance liquid chromatography. Many of the isolates were included in prior taxonomic studies. Sequencing of Nocardia species revealed that members of the group were generally most closely related to the American Type Culture Collection (ATCC) type strains. However, the sequences of Nocardia transvalensis, N. otitidiscaviarum, and N. nova isolates were highly variable; and it is likely that each of these species contains multiple species. We propose that these three species be designated complexes until they are more taxonomically defined. The sequences of several taxa did not match any recognized species. Among other aerobic actinomycetes, each group most closely resembled the associated reference strain, but with some divergence. The study demonstrates the ability of partial 16S rRNA gene sequencing to identify members of the aerobic actinomycetes, but the study also shows that a high degree of sequence divergence exists within many species and that many taxa within the Nocardia spp. are unnamed at present. A major unresolved issue is the type strain of N. asteroides, as the present one (ATCC 19247), chosen before the availability of molecular analysis, does not represent any of the common taxa associated with clinical nocardiosis. PMID:15184431

  6. Combination of low O(2) concentration and mesenchymal stromal cells during culture of cord blood CD34(+) cells improves the maintenance and proliferative capacity of hematopoietic stem cells.

    PubMed

    Hammoud, Mohammad; Vlaski, Marija; Duchez, Pascale; Chevaleyre, Jean; Lafarge, Xavier; Boiron, Jean-Michel; Praloran, Vincent; Brunet De La Grange, Philippe; Ivanovic, Zoran

    2012-06-01

    The physiological approach suggests that an environment associating the mesenchymal stromal cells (MSC) and low O(2) concentration would be most favorable for the maintenance of hematopoietic stem cells (HSCs) in course of ex vivo expansion of hematopoietic grafts. To test this hypothesis, we performed a co-culture of cord blood CD34(+) cells with or without MSC in presence of cytokines for 10 days at 20%, 5%, and 1.5% O(2) and assessed the impact on total cells, CD34(+) cells, committed progenitors (colony-forming cells-CFC) and stem cells activity (pre-CFC and Scid repopulating cells-SRC). Not surprisingly, the expansion of total cells, CD34(+) cells, and CFC was higher in co-culture and at 20% O(2) compared to simple culture and low O(2) concentrations, respectively. However, co-culture at low O(2) concentrations provided CD34(+) cell and CFC amplification similar to classical culture at 20% O(2) . Interestingly, low O(2) concentrations ensured a better pre-CFC and SRC preservation/expansion in co-culture. Indeed, SRC activity in co-culture at 1.5% O(2) was higher than in freshly isolated CD34(+) cells. Interleukin-6 production by MSC at physiologically low O(2) concentrations might be one of the factors mediating this effect. Our data demonstrate that association of co-culture and low O(2) concentration not only induces sufficient expansion of committed progenitors (with respect to the classical culture), but also ensures a better maintenance/expansion of hematopoietic stem cells (HSCs), pointing to the oxygenation as a physiological regulatory factor but also as a cell engineering tool. PMID:21913190

  7. Prevalence of the Most Common Virulence-Associated Genes among Brucella Melitensis Isolates from Human Blood Cultures in Hamadan Province, West of Iran.

    PubMed

    Naseri, Zahra; Alikhani, Mohammad Yousef; Hashemi, Seyed Hamid; Kamarehei, Farideh; Arabestani, Mohammad Reza

    2016-09-01

    Brucellosis is a widespread zoonotic disease causing considerable economic and public health problems. Despite animal vaccination, brucellosis remains endemic in some areas such as Iran, especially in the western Iranian province of Hamadan. We sought to detect some of the most common virulence-associated genes in Brucella isolated from human blood cultures to determine the prevalence of some virulence genes among Brucella isolates. Fifty-seven isolates were studied from patients with a clinical diagnosis of brucellosis who referred to the Infectious Diseases Ward of Sina Hospital in Hamadan Province, Iran, between April 2013 and July 2014. Blood samples were collected for the diagnosis of brucellosis using the BACTEC blood culture system. All of these isolates were confirmed by the bcsp31 Brucella-specific gene. We detected 11 virulence-associated genes of Brucella, namely cβg, virB, znuA, ure, bvfA, omp25, omp31, wbkA, mviN, manA, and manB, which are important for the pathogenesis of this bacterium in the intracellular environment by multiplex PCR. Totally, 149 patients with a clinical diagnosis of brucellosis were enrolled in this study. Fifty-seven (38.3%) patients had positive blood cultures. On biochemical and molecular testing, all of the isolates were Brucella melitensis. Ten of the virulence genes were detected among all of the 57 isolates, but the bvf gene was detected in 53 (93%) isolates. The high prevalence of virulence-associated genes among the Brucella isolates detected in Hamadan Province, Iran, underscores the pathogenicity of this bacterium in this region. PMID:27582592

  8. A Locked Nucleic Acid (LNA)-Based Real-Time PCR Assay for the Rapid Detection of Multiple Bacterial Antibiotic Resistance Genes Directly from Positive Blood Culture

    PubMed Central

    Zhu, Lingxiang; Shen, Dingxia; Zhou, Qiming; Li, Zexia; Fang, Xiangdong; Li, Quan-Zhen

    2015-01-01

    Bacterial strains resistant to various antibiotic drugs are frequently encountered in clinical infections, and the rapid identification of drug-resistant strains is highly essential for clinical treatment. We developed a locked nucleic acid (LNA)-based quantitative real-time PCR (LNA-qPCR) method for the rapid detection of 13 antibiotic resistance genes and successfully used it to distinguish drug-resistant bacterial strains from positive blood culture samples. A sequence-specific primer-probe set was designed, and the specificity of the assays was assessed using 27 ATCC bacterial strains and 77 negative blood culture samples. No cross-reaction was identified among bacterial strains and in negative samples, indicating 100% specificity. The sensitivity of the assays was determined by spiking each bacterial strain into negative blood samples, and the detection limit was 1–10 colony forming units (CFU) per reaction. The LNA-qPCR assays were first applied to 72 clinical bacterial isolates for the identification of known drug resistance genes, and the results were verified by the direct sequencing of PCR products. Finally, the LNA-qPCR assays were used for the detection in 47 positive blood culture samples, 19 of which (40.4%) were positive for antibiotic resistance genes, showing 91.5% consistency with phenotypic susceptibility results. In conclusion, LNA-qPCR is a reliable method for the rapid detection of bacterial antibiotic resistance genes and can be used as a supplement to phenotypic susceptibility testing for the early detection of antimicrobial resistance to allow the selection of appropriate antimicrobial treatment and to prevent the spread of resistant isolates. PMID:25775001

  9. Prevalence of the Most Common Virulence-Associated Genes among Brucella Melitensis Isolates from Human Blood Cultures in Hamadan Province, West of Iran

    PubMed Central

    Naseri, Zahra; Alikhani, Mohammad Yousef; Hashemi, Seyed Hamid; Kamarehei, Farideh; Arabestani, Mohammad Reza

    2016-01-01

    Brucellosis is a widespread zoonotic disease causing considerable economic and public health problems. Despite animal vaccination, brucellosis remains endemic in some areas such as Iran, especially in the western Iranian province of Hamadan. We sought to detect some of the most common virulence-associated genes in Brucella isolated from human blood cultures to determine the prevalence of some virulence genes among Brucella isolates. Fifty-seven isolates were studied from patients with a clinical diagnosis of brucellosis who referred to the Infectious Diseases Ward of Sina Hospital in Hamadan Province, Iran, between April 2013 and July 2014. Blood samples were collected for the diagnosis of brucellosis using the BACTEC blood culture system. All of these isolates were confirmed by the bcsp31 Brucella-specific gene. We detected 11 virulence-associated genes of Brucella, namely cβg, virB, znuA, ure, bvfA, omp25, omp31, wbkA, mviN, manA, and manB, which are important for the pathogenesis of this bacterium in the intracellular environment by multiplex PCR. Totally, 149 patients with a clinical diagnosis of brucellosis were enrolled in this study. Fifty-seven (38.3%) patients had positive blood cultures. On biochemical and molecular testing, all of the isolates were Brucella melitensis. Ten of the virulence genes were detected among all of the 57 isolates, but the bvf gene was detected in 53 (93%) isolates. The high prevalence of virulence-associated genes among the Brucella isolates detected in Hamadan Province, Iran, underscores the pathogenicity of this bacterium in this region. PMID:27582592

  10. Hunting, Swimming, and Worshiping: Human Cultural Practices Illuminate the Blood Meal Sources of Cave Dwelling Chagas Vectors (Triatoma dimidiata) in Guatemala and Belize

    PubMed Central

    Stevens, Lori; Monroy, M. Carlota; Rodas, Antonieta Guadalupe; Dorn, Patricia L.

    2014-01-01

    Background Triatoma dimidiata, currently the major Central American vector of Trypanosoma cruzi, the parasite that causes Chagas disease, inhabits caves throughout the region. This research investigates the possibility that cave dwelling T. dimidiata might transmit the parasite to humans and links the blood meal sources of cave vectors to cultural practices that differ among locations. Methodology/Principal Findings We determined the blood meal sources of twenty-four T. dimidiata collected from two locations in Guatemala and one in Belize where human interactions with the caves differ. Blood meal sources were determined by cloning and sequencing PCR products amplified from DNA extracted from the vector abdomen using primers specific for the vertebrate 12S mitochondrial gene. The blood meal sources were inferred by ≥99% identity with published sequences. We found 70% of cave-collected T. dimidiata positive for human DNA. The vectors had fed on 10 additional vertebrates with a variety of relationships to humans, including companion animal (dog), food animals (pig, sheep/goat), wild animals (duck, two bat, two opossum species) and commensal animals (mouse, rat). Vectors from all locations fed on humans and commensal animals. The blood meal sources differ among locations, as well as the likelihood of feeding on dog and food animals. Vectors from one location were tested for T. cruzi infection, and 30% (3/10) tested positive, including two positive for human blood meals. Conclusions/Significance Cave dwelling Chagas disease vectors feed on humans and commensal animals as well as dog, food animals and wild animals. Blood meal sources were related to human uses of the caves. We caution that just as T. dimidiata in caves may pose an epidemiological risk, there may be other situations where risk is thought to be minimal, but is not. PMID:25211347

  11. Effect of aerobic training and aerobic and resistance training on the inflammatory status of hypertensive older adults.

    PubMed

    Lima, Leandra G; Bonardi, José M T; Campos, Giulliard O; Bertani, Rodrigo F; Scher, Luria M L; Louzada-Junior, Paulo; Moriguti, Júlio C; Ferriolli, Eduardo; Lima, Nereida K C

    2015-08-01

    There is a relationship between high levels of inflammatory markers and low adhesion to the practice of physical activity in the older population. The objective of the present study was to compare the effect of two types of exercise programs, i.e., aerobic training and aerobic plus resistance training on the plasma levels of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) of elderly hypertensive subjects. Hypertensive older volunteers in use of antihypertensive drugs were randomized to three groups: aerobic group (AG), resistance and aerobic group (RAG) and control group (CG). Training lasted 10 weeks, with sessions held three times a week. Blood samples were collected before training and 24 h after completion of the 30 sessions for the determination of serum IL-6 and TNF-α levels. Body mass index was obtained before and after 10 weeks. After intervention, BMI values were lower in AG and RAG compared to CG (p < 0.001), IL-6 was reduced in AG compared to CG (p = 0.04), and TNF-α levels were lower only in RAG compared to CG (p = 0.01). Concluding, both types of training were effective in reducing BMI values in hypertensive older subjects. Aerobic exercise produced the reduction of plasma IL-6 levels. However, the combination of aerobic and resistance exercise, which would be more indicated for the prevention of loss of functionality with aging, showed lower TNF-α mediator after training than control group and a greater fall of TNF-α levels associated to higher BMI reduction. PMID:25567682

  12. The Transition from Aerobic to Anaerobic Metabolism.

    ERIC Educational Resources Information Center

    Skinner, James S.; McLellan, Thomas H.

    1980-01-01

    The transition from aerobic to anaerobic metabolism is discussed. More research is needed on different kinds of athletes and athletic activities and how they may affect aerobic and anaerobic metabolisms. (CJ)

  13. The effect of intensity controlled aerobic dance exercise on aerobic capacity of middle-aged, overweight women.

    PubMed

    Gillett, P A; Eisenman, P A

    1987-12-01

    The purpose of this study was to determine the effect of intensity controlled exercise on the aerobic capacity of overweight, middle-aged women. Thirty-eight moderately overweight women, ages 35-57, participated in a 16-week dance-exercise program. Random assignment was made to an experimental group (n = 20) in which intensity of exercise was controlled and prescribed, and a control group (n = 18) in which exercise was of an intensity typical to commercial aerobic classes. Prior to the onset of training, and at the completion of 16 weeks, the following fitness tests were administered: Aerobic capacity expressed as VO2 max, body composition analysis, blood chemistry, blood pressure, resting heart rate, muscular endurance, and flexibility. T-tests, ANCOVA, and gain-score analyses were utilized to evaluate data. Both groups showed small changes in weight, percent fat, resting systolic and diastolic blood pressure, resting heart rate, high density lipoprotein-cholesterol (HDL-C), muscular endurance, and flexibility, but these changes were statistically nonsignificant. The VO2 max for the experimental group increased 41%, while the VO2 max for the control group increased 22% (p less than 0.05). The results suggest that the cardiovascular fitness changes for overweight, middle-aged women are greater when exercise intensity and progression are tailored to their age and fitness level. PMID:3423310

  14. Aerobic Microbial Degradation of Glucoisosaccharinic Acid

    PubMed Central

    Strand, S. E.; Dykes, J.; Chiang, V.

    1984-01-01

    α-Glucoisosaccharinic acid (GISA), a major by-product of kraft paper manufacture, was synthesized from lactose and used as the carbon source for microbial media. Ten strains of aerobic bacteria capable of growth on GISA were isolated from kraft pulp mill environments. The highest growth yields were obtained with Ancylobacter spp. at pH 7.2 to 9.5. GISA was completely degraded by cultures of an Ancylobacter isolate. Ancylobacter cell suspensions consumed oxygen and produced carbon dioxide in response to GISA addition. A total of 22 laboratory strains of bacteria were tested, and none was capable of growth on GISA. GISA-degrading isolates were not found in forest soils. Images PMID:16346467

  15. New Routes for Aerobic Biodegradation of Dimethylsulfoniopropionate

    PubMed Central

    Taylor, Barrie F.; Gilchrist, Darrin C.

    1991-01-01

    Dimethylsulfoniopropionate (DMSP), an osmolyte in marine plants, is biodegraded by cleavage of dimethyl sulfide (DMS) or by demethylation to 3-methiolpropionate (MMPA) and 3-mercaptopropionate (MPA). Sequential demethylation has been observed only with anoxic slurries of coastal sediments. Bacteria that grew aerobically on MMPA and DMSP were isolated from marine environments and phytoplankton cultures. Enrichments with DMSP selected for bacteria that generated DMS, whereas MMPA enrichments selected organisms that produced methanethiol (CH3SH) from either DMSP or MMPA. A bacterium isolated on MMPA grew on MMPA and DMSP, but rapid production of CH3SH from DMSP occurred only with DMSP-grown cells. Low levels of MPA accumulated during growth on MMPA, indicating demethylation as well as demethiolation of MMPA. The alternative routes for DMSP biodegradation via MMPA probably impact on net DMS fluxes to the marine atmosphere. PMID:16348607

  16. Anaerobic/aerobic treatment of coloured textile effluents using sequencing batch reactors.

    PubMed

    Shaw, C B; Carliell, C M; Wheatley, A D

    2002-04-01

    Conventional biological wastewater treatment plants do not easily degrade the dyes and polyvinyl alcohols (PVOH) in textile effluents. Results are reported on the possible advantages of anaerobic/aerobic cometabolism in sequenced redox reactors. A six phase anaerobic/aerobic sequencing laboratory scale batch reactor was developed to treat a synthetic textile effluent. The wastewater included PVOH from desizing and an azo dye (Remazol Black). The reactor removed 66% of the applied total organic carbon (load F: M 0.15) compared to 76% from a control reactor without dye. Colour removal was 94% but dye metabolites caused reactor instability. Aromatic amines from the anaerobic breakdown of the azo dyes were not completely mineralised by the aerobic phase. Breakdown of PVOH by the reactor (20-30%) was not as good as previous reports with entirely aerobic cultures. The anaerobic cultures were able to tolerate the oxygen and methane continued to be produced but there was a deterioration in settlement. PMID:12092574

  17. Regulation of IgA secretion in polyclonally induced in vitro human lymphocyte cultures: the function of T and B cells from mesenteric lymph nodes and peripheral blood.

    PubMed Central

    Pang, G; Yeung, S; Clancy, R L; Cripps, A W; Hennessy, E J; Santhanam, A N

    1986-01-01

    Human gut-associated immunoregulatory events were studied in a pokeweed mitogen (PWM)-stimulated culture system using lymphocytes obtained from the mesenteric lymph nodes (MLN) of female subjects undergoing gastroplasty for obesity. Compared with peripheral blood lymphocytes, lymphocytes obtained from MLN secreted IgG, IgA and IgM isotypes that differ in pattern and distribution despite similar proportions of T cells and B cells expressing isotype-specific surface membrane immunoglobulin (SmIg). Among the isotypes secreted, IgA appeared to be increased relatively to other isotypes in MLN cultures. Crossover coculture experiments using T and B cells isolated from both MLN and blood by E-rosetting and cell panning procedures demonstrated that IgA was particularly sensitive to help and suppression exerted by MLN T cells and T cell subsets defined by monoclonal antibodies OKT4 and OKT8 respectively, when compared with similar subsets isolated from blood. The results presented provide a basis for study of gut handling of ingested antigen in man, and of disturbed immunoregulatory events in inflammatory and neoplastic disease of the human gut. PMID:2942320

  18. PRC2 inhibition counteracts the culture-associated loss of engraftment potential of human cord blood-derived hematopoietic stem and progenitor cells

    PubMed Central

    Varagnolo, Linda; Lin, Qiong; Obier, Nadine; Plass, Christoph; Dietl, Johannes; Zenke, Martin; Claus, Rainer; Müller, Albrecht M.

    2015-01-01

    Cord blood hematopoietic stem cells (CB-HSCs) are an outstanding source for transplantation approaches. However, the amount of cells per donor is limited and culture expansion of CB-HSCs is accompanied by a loss of engraftment potential. In order to analyze the molecular mechanisms leading to this impaired potential we profiled global and local epigenotypes during the expansion of human CB hematopoietic stem and progenitor cells (HPSCs). Human CB-derived CD34+ cells were cultured in serum-free medium together with SCF, TPO, FGF, with or without Igfbp2 and Angptl5 (STF/STFIA cocktails). As compared to the STF cocktail, the STFIA cocktail maintains in vivo repopulation capacity of cultured CD34+ cells. Upon expansion, CD34+ cells genome-wide remodel their epigenotype and depending on the cytokine cocktail, cells show different H3K4me3 and H3K27me3 levels. Expanding cells without Igfbp2 and Angptl5 leads to higher global H3K27me3 levels. ChIPseq analyses reveal a cytokine cocktail-dependent redistribution of H3K27me3 profiles. Inhibition of the PRC2 component EZH2 counteracts the culture-associated loss of NOD scid gamma (NSG) engraftment potential. Collectively, our data reveal chromatin dynamics that underlie the culture-associated loss of engraftment potential. We identify PRC2 component EZH2 as being involved in the loss of engraftment potential during the in vitro expansion of HPSCs. PMID:26198814

  19. Rapid Identification of Bacteria Directly from Positive Blood Cultures by Use of a Serum Separator Tube, Smudge Plate Preparation, and Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry.

    PubMed

    Chen, Yan; Porter, Vanessa; Mubareka, Samira; Kotowich, Leona; Simor, Andrew E

    2015-10-01

    We analyzed the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) of smudge plate growth for bacterial identification from 400 blood cultures. Ninety-seven percent of Gram-negative bacilli and 85% of Gram-positive organisms were correctly identified within 4 h; only eight isolates (2.0%) were misidentified. This method provided rapid and accurate microbial identification from positive blood cultures. PMID:26202115

  20. Moving blood.

    PubMed

    Pelis, K

    1997-01-01

    Our internationally acclaimed journalist Sanguinia has returned safely from her historic assignment. Travelling from Homeric Greece to British Romanticism, she was witness to blood drinking, letting, bathing, and transfusion. In this report, she explores connections between the symbolic and the sadistic; the mythic and the medical--all in an effort to appreciate the layered meanings our culture has given to the movement of blood between our bodies. PMID:9407636

  1. Pediatric Multicenter Evaluation of the Verigene Gram-Negative Blood Culture Test for Rapid Detection of Inpatient Bacteremia Involving Gram-Negative Organisms, Extended-Spectrum Beta-Lactamases, and Carbapenemases

    PubMed Central

    Deburger, B.; Roundtree, S. S.; Ventrola, C. A.; Blecker-Shelly, D. L.; Mortensen, J. E.

    2014-01-01

    We evaluated the investigational use only (IUO) version of the rapid Verigene Gram-negative blood culture test (BC-GN), a microarray that detects 9 genus/species targets (Acinetobacter spp., Citrobacter spp., Enterobacter spp., Escherichia coli/Shigella spp., Klebsiella oxytoca, Klebsiella pneumoniae, Proteus spp., Pseudomonas aeruginosa, and Serratia marcescens) and 6 antimicrobial resistance determinants (blaCTX-M, blaKPC, blaNDM, blaVIM, blaIMP, and blaOXA) directly from positive blood cultures. BC-GN was performed on positive BacT/Alert Pediatric FAN and Bactec Peds Plus blood cultures with Gram-negative organisms at two tertiary pediatric centers. Vitek MS (bioMérieux, Durham, NC) was used to assign gold standard organism identification. The Check MDR CT-102 microarray (Check Points B.V., Wageningen, Netherlands) was used as an alternative method for detecting resistance determinants. In total, 104 organisms were isolated from 97 clinical blood cultures. BC-GN correctly detected 26/26 cultures with Acinetobacter spp., P. aeruginosa, and S. marcescens, 5/6 with Citrobacter spp., 13/14 with Enterobacter spp., 23/24 with E. coli, 2/3 with K. oxytoca, 16/17 with K. pneumoniae, and 0/1 with Proteus spp. BC-GN appropriately reported negative BC-GN results in 8/13 blood cultures that grew organisms that were not represented on the microarray but failed to detect targets in 3/5 cultures that grew multiple Gram-negative organisms. BC-GN detected 5/5 and 1/1 clinical blood cultures with blaCTX-M and blaVIM. All 6 results were corroborated by Check MDR CT-102 microarray testing. The Verigene BC-GN test has the potential to expedite therapeutic decision making in pediatric patients with Gram-negative bacteremia. Sensitivity was satisfactory but may be suboptimal in mixed Gram-negative blood cultures. PMID:24759724

  2. "Aerobic" Writing: A Writing Practice Model.

    ERIC Educational Resources Information Center

    Crisp, Sally Chandler

    "Aerobic writing" is a writing center strategy designed to keep students in writing "shape." Like aerobic exercise, aerobic writing is sustained for a certain length of time and done on a regular basis at prescribed time intervals. The program requires students to write at least two times a week for approximately an hour each time. Students write,…

  3. Arthritis and Aerobic Exercise: A Review.

    ERIC Educational Resources Information Center

    Ike, Robert W.; And Others

    1989-01-01

    Arthritic patients who regularly do aerobic exercise make significant gains in aerobic and functional status, and in subjective areas like pain tolerance and mood. Still, they are often advised to curtail physical activity. Guidelines are presented for physicians prescribing aerobic exercise. An exercise tolerance test is recommended. (SM)

  4. Analysis of Aerobic Respiration in Intact Skeletal Muscle Tissue by Microplate-Based Respirometry.

    PubMed

    Shintaku, Jonathan; Guttridge, Denis C

    2016-01-01

    Mitochondrial function is a key component of skeletal muscle health, and its dysfunction has been associated with a wide variety of diseases. Microplate-based respirometry measures aerobic respiration of live cells through extracellular changes in oxygen concentration. Here, we describe a methodology to measure aerobic respiration of intact murine skeletal muscle tissue. The tissues are not cultured, permeabilized, or enzymatically dissociated to single fibers, so there is minimal experimental manipulation affecting the samples prior to acquiring measurements. PMID:27492183

  5. [Investigation of plasmid mediated AmpC beta-lactamases among Escherichia coli and Klebsiella pneumoniae isolated from blood cultures].

    PubMed

    Sarı, Ayşe Nur; Biçmen, Meral; Gülay, Zeynep

    2013-10-01

    The aim of this study was to investigate the prevalence and types of plasmid-mediated AmpC (pAmpC) beta-lactamase enzymes in Escherichia coli and Klebsiella pneumoniae strains isolated from blood cultures of hospitalized patients in Dokuz Eylul University Hospital between 2007 and 2012. A total of 261 isolates which consisted of 184 E.coli (70.5%) and 77 K.pneumoniae (29.5%) were included in the study. All isolates were resistant to cefotaxime and/or ceftazidime but susceptible to imipenem. Cefoxitin resistance was investigated as an indicator of AmpC type enzymes. A total of 57 (21.8%) isolates which were cefoxitin-resistant (32 E.coli, 25 K.pneumoniae), were screened for pampC genes by a multiplex polymerase chain reaction (PCR) assay. Additionally, 10 of each cefoxitin susceptible isolates per year were chosen randomly and screened by the same PCR assay to detect the presence of ACC enzymes, which can not hydrolyze cefoxitin. Positive PCR results were confirmed by sequence analysis. Plasmid analysis and macrorestriction analysis were performed for pampC-positive isolates. The presence of pAmpC enzymes has been shown in 9.4% (3/32) of cefoxitin-resistant E.coli, and 8% (2/25) of cefoxitin-resistant K.pneumoniae strains. It was noted that there were no strains producing this enzyme isolated in 2007 and 2008, however the prevalence of pAmpC was detected as 1.6% in 2009 (one ACT-1 producing K.pneumoniae), increasing to 4.8% in 2011 (one ACT-1 producing K.pneumoniae) and 6.4% in 2012 (three CMY-2 producing E.coli). These enzymes were found to be carried on 81 kb size plasmids in K.pneumoniae isolates and on a 9 kb size plasmid in E.coli isolates. Macrorestriction analysis indicated that two of the three CMY-2 producing E.coli had the same PFGE (Pulsed-field gel electrophoresis) pattern. If these two strains are considered as identical, it can be concluded that the prevalence of pAmpC was low in the strains isolated between 2007-2012 (4/261; 1.5%) in our institution

  6. In Vitro Culture During Retroviral Transduction Improves Thymic Repopulation and Output After Total Body Irradiation and Autologous Peripheral Blood Progenitor Cell Transplantation in Rhesus Macaques

    PubMed Central

    Loré, Karin; Seggewiss, Ruth; Guenaga, F. Javier; Pittaluga, Stefania; Donahue, Robert E.; Krouse, Allen; Metzger, Mark E.; Koup, Richard A.; Reilly, Cavan; Douek, Daniel C.; Dunbar, Cynthia E.

    2008-01-01

    Immunodeficiency after peripheral blood progenitor cell (PBPC) transplantation may be influenced by graft composition, underlying disease, and/or pre-treatment. These factors are difficult to study independently in humans. Ex vivo culture and genetic manipulation of PBPC grafts may also affect immune reconstitution, with relevance to gene therapy applications. We directly compared the effects of three clinically relevant autologous graft compositions on immune reconstitution after myeloblative total body irradiation in rhesus macaques, the first time these studies have been performed in a large animal model with direct clinical relevance. Animals received CD34+ cell dose-matched grafts of either peripheral blood mononuclear cells, purified CD34+ PBPCs, or purified CD34+ PBPCs expanded in vitro and retrovirally transduced. We evaluated the reconstitution of T, B, natural killer, dendritic cells, and monocytes in blood and lymph nodes for up to 1 year post-transplantation. Animals receiving selected-transduced CD34+ cells had the fastest recovery of T-cell numbers, along with the highest T-cell-receptor gene rearrangement excision circles levels, the fewest proliferating Ki-67+ T-cells in the blood, and the best-preserved thymic architecture. Selected-transduced CD34+ cells may therefore repopulate the thymus more efficiently and promote a higher output of naïve T-cells. These results have implications for the design of gene therapy trials, as well as for the use of expanded PBPCs for improved T-cell immune reconstitution after transplantation. PMID:16497945

  7. Molecular characterization of Italian Candida parapsilosis isolates reveals the cryptic presence of the newly described species Candida orthopsilosis in blood cultures from newborns.

    PubMed

    Romeo, Orazio; Delfino, Demetrio; Costanzo, Barbara; Cascio, Antonio; Criseo, Giuseppe

    2012-03-01

    The authors report the molecular characterization of Candida parapsilosis isolates recovered from the blood and venous central catheter tips of patients admitted to different care units of the Polyclinic Hospital, University of Messina, Italy. Among 97 presumed C. parapsilosis isolates examined, 94 were identified as C. parapsilosis sensu stricto and the remaining 3 isolates were found to belong to the cryptic species Candida orthopsilosis which was recovered only from blood cultures of neonates (<30 days old) born prematurely. No C. metapsilosis was found in this study. This study emphasizes the role of C. parapsilosis as an important nosocomial pathogen, and it also describes, for the first time, the occurrence of C. orthopsilosis in newborns. PMID:22244186

  8. Molecular epidemiology and antimicrobial susceptibility profiles of methicillin-resistant Staphylococcus aureus blood culture isolates: results of the Quebec Provincial Surveillance Programme.

    PubMed

    Lévesque, S; Bourgault, A M; Galarneau, L A; Moisan, D; Doualla-Bell, F; Tremblay, C

    2015-05-01

    The objectives of this study were to characterize methicillin-resistant Staphylococcus aureus (MRSA) blood culture isolates and to determine their relative importance in both nosocomial and community-acquired infections. A total of 535 MRSA blood culture isolates were analysed. In vitro susceptibility to 14 agents was determined. The genes nuc, mecA and coding for PVL toxin were identified by PCR. All isolates were characterized by PFGE or spa typing to assess their genomic relationships. Most MRSA isolates were retrieved from nosocomial bloodstream infections (474, 89%) and were of the CMRSA2 genotype. Healthcare-associated (HA)-MRSA bloodstream infections were associated with older age (70-89 years, P = 0·002) and most often secondary to central line infections (P = 0·005). Among MRSA strains associated with community-acquired (CA)-MRSA, 28·8% were isolated in intravenous drug users. CA-MRSA genotypes were more frequently found in young adults (20-39 years, P < 0·0001) with skin/soft tissue as the primary sources of infection (P = 0·006). CMRSA10 genotype was the predominant CA-MRSA strain. All MRSA isolates were susceptible to doxycycline, tigecycline, trimethoprim/sulfamethoxazole and vancomycin. Both the presence of the genes coding for PVL toxin (89·8%) and susceptibility to clindamycin (86·5%) were predictive of CA-MRSA genotypes. Whereas in the USA, HA-MRSA have been replaced by USA300 (CMRSA10) clone as the predominant MRSA strain type in positive blood cultures from hospitalized patients, this phenomenon has not been observed in the province of Quebec. PMID:25140694

  9. Comparison of the Staphylococcus QuickFISH BC test with the tube coagulase test performed on positive blood cultures for evaluation and application in a clinical routine setting.

    PubMed

    Carretto, E; Bardaro, M; Russello, G; Mirra, M; Zuelli, C; Barbarini, D

    2013-01-01

    Many studies demonstrate that delayed proper therapy in bloodstream infections caused by Staphylococcus aureus increases the mortality rate, emphasizing the need to shorten the turnaround time for positive blood cultures. Different techniques are currently available, from phenotypic methods to more complex tests such as matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF), real-time PCR (RT-PCR), and fluorescence in situ hybridization using peptide nucleic acid probes (PNA FISH). This study evaluated the performance of the Staphylococcus QuickFISH BC test (QFT), a novel FISH methodology, compared with the direct tube coagulase test (DTCT) on blood cultures exhibiting Gram-positive cocci in clusters. A total of 173 blood cultures collected from 128 different patients were analyzed using the DTCT, evaluated after both 4 and 24 h, and the QFT. A total of 179 isolates were identified using the Vitek2 system. Thirty-five out of 35 Staphylococcus aureus were correctly identified by the QFT (sensitivity = 100%), with a specificity of 100% (no green fluorescence was detected for strains different from S. aureus). The DTCT was positive after 4 h for 28 out of the 35 samples (sensitivity = 80%) and after 24 h for 31 out of the 35 samples (sensitivity = 88.57%). Among the remaining 144 isolates, one was then identified as Corynebacterium striatum and two as Micrococcus luteus. QFT identified 139 out of the 141 coagulase-negative staphylococci (CoNS) (sensitivity = 98.58%), showing again a specificity of 100% (no fluorescent red signals were detected for strains different from CoNS). We also discuss also the implementation process of this methodology in our setting, with particular emphasis on the workflow and the cost-effectiveness. PMID:23100336

  10. Use of Disinfection Cap to Reduce Central-Line–Associated Bloodstream Infection and Blood Culture Contamination Among Hematology–Oncology Patients

    PubMed Central

    Kamboj, Mini; Blair, Rachel; Bell, Natalie; Son, Crystal; Huang, Yao-Ting; Dowling, Mary; Lipitz-Snyderman, Allison; Eagan, Janet; Sepkowitz, Kent

    2016-01-01

    OBJECTIVE In this study, we examined the impact of routine use of a passive disinfection cap for catheter hub decontamination in hematology–oncology patients. SETTING A tertiary care cancer center in New York City METHODS In this multiphase prospective study, we used 2 preintervention phases (P1 and P2) to establish surveillance and baseline rates followed by sequential introduction of disinfection caps on high-risk units (HRUs: hematologic malignancy wards, hematopoietic stem cell transplant units and intensive care units) (P3) and general oncology units (P4). Unit-specific and hospital-wide hospital-acquired central-line–associated bloodstream infection (HA-CLABSI) rates and blood culture contamination (BCC) with coagulase negative staphylococci (CONS) were measured. RESULTS Implementation of a passive disinfection cap resulted in a 34% decrease in hospital-wide HA-CLABSI rates (combined P1 and P2 baseline rate of 2.66–1.75 per 1,000 catheter days at the end of the study period). This reduction occurred only among high-risk patients and not among general oncology patients. In addition, the use of the passive disinfection cap resulted in decreases of 63% (HRUs) and 51% (general oncology units) in blood culture contamination, with an estimated reduction of 242 BCCs with CONS. The reductions in HA-CLABSI and BCC correspond to an estimated annual savings of $3.2 million in direct medical costs. CONCLUSION Routine use of disinfection caps is associated with decreased HA-CLABSI rates among high-risk hematology oncology patients and a reduction in blood culture contamination among all oncology patients. PMID:26394849

  11. 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.

    PubMed

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

    2016-06-01

    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

  12. Comparison of Coagulase-Negative Staphylococci Isolated from Blood Cultures as a True Bacteremia Agent and Contaminant in Terms of Slime Production and Methicillin Resistance

    PubMed Central

    Uyanik, Muhammet Hamidullah; Yazgi, Halil; Ozden, Kemalettin; Erdil, Zeynep; Ayyildiz, Ahmet

    2014-01-01

    Objective: The aim of this study is to determine the species distribution, slime activity, and methicillin resistance of coagulase-negative staphylococci (CoNS) isolated from blood cultures as either contaminants or true bacteremia agents. Materials and Methods: In this study, 13.268 blood culture samples sent to our laboratory from various clinics during a two-year period were examined in terms of the presence of CoNS to clarify whether the isolates are true bacteremia agents, as defined by Centers for Disease Control and Prevention (CDC) criteria. The slime activities of true bacteremia agents (58 CoNS strains) and contaminants (50 randomly selected CoNS strains) were investigated by the Christensen method. The methicillin susceptibilities of the strains were determined by the disk diffusion method. Results: Although the frequency of slime production was 39.7% among the true bacteremia CoNS agents, it was 18% in CoNS that were judged to be contaminants (p<0.05). S. epidermidis was the most frequently isolated species for both the true bacteremia agent group (56.9%) and contaminant group (74%). Additionally, S. epidermidis was the bacterium most frequently characterized as slime producing in both groups. The methicillin resistance of slime-producing CoNS was determined to be 82.6% for the true bacteremia agent group and 77.8% for the contaminant group. Conclusion: The presence of slime activity in CoNS isolated from blood culture samples is supportive evidence that they are most likely the agents of true bacteremia cases. PMID:25610309

  13. Rapid Testing Using the Verigene Gram-Negative Blood Culture Nucleic Acid Test in Combination with Antimicrobial Stewardship Intervention against Gram-Negative Bacteremia

    PubMed Central

    Leekha, Surbhi; Heil, Emily L.; Zhao, LiCheng; Badamas, Rilwan; Johnson, J. Kristie

    2014-01-01

    Rapid identification of microorganisms and antimicrobial resistance is paramount for targeted treatment in serious bloodstream infections (BSI). The Verigene Gram-negative blood culture nucleic acid test (BC-GN) is a multiplex, automated molecular diagnostic test for identification of eight Gram-negative (GN) organisms and resistance markers from blood culture with a turnaround time of approximately 2 h. Clinical isolates from adult patients at the University Maryland Medical Center with GN bacteremia from 1 January 2012 to 30 June 2012 were included in this study. Blood culture bottles were spiked with clinical isolates, allowed to incubate, and processed by BC-GN. A diagnostic evaluation was performed. In addition, a theoretical evaluation of time to effective and optimal antibiotic was performed, comparing actual antibiotic administration times from chart review (“control”) to theoretical administration times based on BC-GN reporting and antimicrobial stewardship team (AST) review (“intervention”). For organisms detected by the assay, BC-GN correctly identified 95.6% (131/137), with a sensitivity of 97.1% (95% confidence interval [CI], 90.7 to 98.4%) and a specificity of 99.5% (95% CI, 98.8 to 99.8%). CTX-M and OXA resistance determinants were both detected. Allowing 12 h from Gram stain for antibiotic implementation, the intervention group had a significantly shorter duration to both effective (3.3 versus 7.0 h; P < 0.01) and optimal (23.5 versus 41.8 h; P < 0.01) antibiotic therapy. BC-GN with AST intervention can potentially decrease time to both effective and optimal antibiotic therapy in GN BSI. PMID:25547353

  14. Effects of Aerobic Exercise Based upon Heart Rate at Aerobic Threshold in Obese Elderly Subjects with Type 2 Diabetes

    PubMed Central

    Donini, Lorenzo Maria

    2015-01-01

    In obese diabetic subjects, a correct life style, including diet and physical activity, is part of a correct intervention protocol. Thus, the aim of this study was to evaluate the effects of aerobic training intervention, based on heart rate at aerobic gas exchange threshold (AerTge), on clinical and physiological parameters in obese elderly subjects with type 2 diabetes (OT2DM). Thirty OT2DM subjects were randomly assigned to an intervention (IG) or control group (CG). The IG performed a supervised aerobic exercise training based on heart rate at AerTge whereas CG maintained their usual lifestyle. Anthropometric measures, blood analysis, peak oxygen consumption (V˙O2peak), metabolic equivalent (METpeak), work rate (WRpeak), and WRAerTge were assessed at baseline and after intervention. After training, patients enrolled in the IG had significantly higher (P < 0.001) V˙O2peak, METpeak, WRpeak, and WRAerTge and significantly lower (P < 0.005) weight, BMI, %FM, and waist circumference than before intervention. Both IG and CG subjects had lower glycated haemoglobin levels after intervention period. No significant differences were found for all the other parameters between pre- and posttraining and between groups. Aerobic exercise prescription based upon HR at AerTge could be a valuable physical intervention tool to improve the fitness level and metabolic equilibrium in OT2DM patients. PMID:26089890

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

    PubMed

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

    2013-04-01

    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

  16. Clinical-Grade Generation of Active NK Cells from Cord Blood Hematopoietic Progenitor Cells for Immunotherapy Using a Closed-System Culture Process

    PubMed Central

    Spanholtz, Jan; Preijers, Frank; Tordoir, Marleen; Trilsbeek, Carel; Paardekooper, Jos; de Witte, Theo; Schaap, Nicolaas; Dolstra, Harry

    2011-01-01

    Natural killer (NK) cell-based adoptive immunotherapy is a promising treatment approach for many cancers. However, development of protocols that provide large numbers of functional NK cells produced under GMP conditions are required to facilitate clinical studies. In this study, we translated our cytokine-based culture protocol for ex vivo expansion of NK cells from umbilical cord blood (UCB) hematopoietic stem cells into a fully closed, large-scale, cell culture bioprocess. We optimized enrichment of CD34+ cells from cryopreserved UCB units using the CliniMACS system followed by efficient expansion for 14 days in gas-permeable cell culture bags. Thereafter, expanded CD34+ UCB cells could be reproducibly amplified and differentiated into CD56+CD3− NK cell products using bioreactors with a mean expansion of more than 2,000 fold and a purity of >90%. Moreover, expansion in the bioreactor yielded a clinically relevant dose of NK cells (mean: 2×109 NK cells), which display high expression of activating NK receptors and cytolytic activity against K562. Finally, we established a versatile closed washing procedure resulting in optimal reduction of medium, serum and cytokines used in the cell culture process without changes in phenotype and cytotoxic activity. These results demonstrate that large numbers of UCB stem cell-derived NK cell products for adoptive immunotherapy can be produced in closed, large-scale bioreactors for the use in clinical trials. PMID:21698239

  17. Obesity promotes aerobic glycolysis in prostate cancer cells.

    PubMed

    Cavazos, David A; deGraffenried, Matthew J; Apte, Shruti A; Bowers, Laura W; Whelan, Kaitlin A; deGraffenried, Linda A

    2014-01-01

    Obesity is the leading preventable comorbidity associated with increased prostate cancer-related recurrence and mortality. Epidemiological and clinical studies indicate that a body mass index >30 is associated with increased oxidative DNA damage within the prostate gland and increased prostate cancer-related mortality. Here we provide evidence that obesity promotes worse clinical outcome through induction of metabolic abnormalities known to promote genotoxic stress. We have previously reported that blood serum derived from obese mice may enhance the proliferative and invasive potential of human prostate cancer cell lines ex vivo. Here we show that a 1-h exposure of LNCaP or PacMetUT1 prostate cancer cell lines and nonmalignant RWPE-1 prostate epithelial cells to 2% serum from obese mice induces markers of aerobic glycolysis relative to those exposed to serum from nonobese mice. This metabolic change was correlated with accumulation of reactive oxygen species (ROS) and increased frequency of DNA double-strand breaks. Interestingly, N-tert-Butylhydroxylamine, an antioxidant, significantly suppressed markers of aerobic glycolysis in the cells exposed to the blood serum of obese mice, suggesting that ROS contributes to a metabolic shift toward aerobic glycolysis. Here we describe obesity-induced changes in key metabolic markers that impact prostate cancer cell progression and explore the role of antioxidants in ameliorating these effects. PMID:25264717

  18. Biodegradation of Asphalt Cement-20 by Aerobic Bacteria

    PubMed Central

    Pendrys, John P.

    1989-01-01

    Seven gram-negative, aerobic bacteria were isolated from a mixed culture enriched for asphalt-degrading bacteria. The predominant genera of these isolates were Pseudomonas, Acinetobacter, Alcaligenes, Flavimonas, and Flavobacterium. The mixed culture preferentially degraded the saturate and naphthene aromatic fractions of asphalt cement-20. A residue remained on the surface which was resistant to biodegradation and protected the underlying asphalt from biodegradation. The most potent asphalt-degrading bacterium, Acinetobacter calcoaceticus NAV2, excretes an emulsifier which is capable of emulsifying the saturate and naphthene aromatic fractions of asphalt cement-20. This emulsifier is not denatured by phenol. PMID:16347928

  19. Blood-Compatible Polymer for Hepatocyte Culture with High Hepatocyte-Specific Functions toward Bioartificial Liver Development.

    PubMed

    Hoshiba, Takashi; Otaki, Takayuki; Nemoto, Eri; Maruyama, Hiroka; Tanaka, Masaru

    2015-08-19

    The development of bioartificial liver (BAL) is expected because of the shortage of donor liver for transplantation. The substrates for BAL require the following criteria: (a) blood compatibility, (b) hepatocyte adhesiveness, and (c) the ability to maintain hepatocyte-specific functions. Here, we examined blood-compatible poly(2-methoxyethyl acrylate) (PMEA) and poly(tetrahydrofurfuryl acrylate) (PTHFA) (PTHFA) as the substrates for BAL. HepG2, a human hepatocyte model, could adhere on PMEA and PTHFA substrates. The spreading of HepG2 cells was suppressed on PMEA substrates because integrin contribution to cell adhesion on PMEA substrate was low and integrin signaling was not sufficiently activated. Hepatocyte-specific gene expression in HepG2 cells increased on PMEA substrate, whereas the expression decreased on PTHFA substrates due to the nuclear localization of Yes-associated protein (YAP). These results indicate that blood-compatible PMEA is suitable for BAL substrate. Also, PMEA is expected to be used to regulate cell functions for blood-contacting tissue engineering. PMID:26258689

  20. Temporal expression of transporters and receptors in a rat primary co-culture blood-brain barrier model.

    PubMed

    Liu, Houfu; Li, Yang; Lu, Sijie; Wu, Yiwen; Sahi, Jasminder

    2014-10-01

    1. The more relevant primary co-cultures of brain microvessel endothelial cells and astrocytes (BMEC) are less utilized for screening of potential CNS uptake when compared to intestinal and renal cell lines. 2. In this study, we characterized the temporal mRNA expression of major CNS transporters and receptors, including the transporter regulators Pxr, Ahr and Car in a rat BMEC co-cultured model. Permeability was compared with the Madin-Darby canine kidney (MDCKII)-MDR1 cell line and rat brain in situ perfusion model. 3. Our data demonstrated differential changes in expression of individual transporters and receptors over the culture period. Expression of ATP-binding cassette transporters was better retained than that of solute carrier transporters. The insulin receptor (IR) was best maintained among investigated receptors. AhR demonstrated high mRNA expression in rat brain capillaries and expression was better retained than Pxr or Car in culture. Mdr1b expression was up-regulated during primary culture, albeit Mdr1a mRNA levels were much higher. P-gp and Bcrp-1 were highly expressed and functional in this in vitro system. 4. Permeability measurements with 18 CNS marketed drugs demonstrated weak correlation between rBMEC model and rat in situ permeability and moderate correlation with MDCKII-MDR1 cells. 5. We have provided appropriate methodologies, as well as detailed and quantitative characterization data to facilitate improved understanding and rational use of this in vitro rat BBB model. PMID:24827375

  1. Aerobic microbial enhanced oil recovery

    SciTech Connect

    Torsvik, T.; Gilje, E.; Sunde, E.

    1995-12-31

    In aerobic MEOR, the ability of oil-degrading bacteria to mobilize oil is used to increase oil recovery. In this process, oxygen and mineral nutrients are injected into the oil reservoir in order to stimulate growth of aerobic oil-degrading bacteria in the reservoir. Experiments carried out in a model sandstone with stock tank oil and bacteria isolated from offshore wells showed that residual oil saturation was lowered from 27% to 3%. The process was time dependent, not pore volume dependent. During MEOR flooding, the relative permeability of water was lowered. Oxygen and active bacteria were needed for the process to take place. Maximum efficiency was reached at low oxygen concentrations, approximately 1 mg O{sub 2}/liter.

  2. Peripheral Blood Monocytes as Adult Stem Cells: Molecular Characterization and Improvements in Culture Conditions to Enhance Stem Cell Features and Proliferative Potential

    PubMed Central

    Ungefroren, Hendrik; Hyder, Ayman; Schulze, Maren; Fawzy El-Sayed, Karim M.; Grage-Griebenow, Evelin; Nussler, Andreas K.; Fändrich, Fred

    2016-01-01

    Adult stem or programmable cells hold great promise in diseases in which damaged or nonfunctional cells need to be replaced. We have recently demonstrated that peripheral blood monocytes can be differentiated in vitro into cells resembling specialized cell types like hepatocytes and pancreatic beta cells. During phenotypic conversion, the monocytes downregulate monocyte/macrophage differentiation markers, being indicative of partial dedifferentiation, and are partially reprogrammed to acquire a state of plasticity along with expression of various markers of pluripotency and resumption of mitosis. Upregulation of stem cell markers and mitotic activity in the cultures was shown to be controlled by autocrine production/secretion of activin A and transforming growth factor-beta (TGF-β). These reprogrammed monocyte derivatives were termed “programmable cells of monocytic origin” (PCMO). Current efforts focus on establishing culture conditions that increase both the plasticity and proliferation potential of PCMO in order to be able to generate large amounts of blood-derived cells suitable for both autologous and allogeneic therapies. PMID:26798361

  3. WWOX loss activates aerobic glycolysis

    PubMed Central

    Abu-Remaileh, Muhannad; Seewaldt, Victoria L; Aqeilan, Rami I

    2015-01-01

    Cancer cells undergo reprogramming of glucose metabolism to limit energy production to glycolysis—a state known as “aerobic glycolysis.” Hypoxia-inducible factor 1 (HIF1α) is a transcription factor that regulates many genes responsible for this switch. As discussed here, new data suggest that the tumor suppressor WW domain-containing oxidoreductase (WWOX) modulates HIF1α, thereby regulating this metabolic state. PMID:27308416

  4. Aerobic and Anaerobic Bacteriology of Hidradenitis Suppurativa: A Study of 22 Cases

    PubMed Central

    Katoulis, Alexandros C.; Koumaki, Dimitra; Liakou, Aikaterini I.; Vrioni, Georgia; Koumaki, Vasiliki; Kontogiorgi, Dimitra; Tzima, Korina; Tsakris, Athanasios; Rigopoulos, Dimitris

    2015-01-01

    Introduction Hidradenitis suppurativa (HS) is a chronic inflammatory skin disease of unclear etiology. The role of bacteria in the pathogenesis of disease remains controversial. Materials and Methods Specimens were obtained from 22 HS patients by direct percutaneous needle aspiration. The collected material was cultured in aerobic and anaerobic conditions, and sensitivity tests were performed. Results Of the 22 patients, 32% were culture negative and 68% were culture positive. A total of 16 isolates was obtained, 14 aerobic and 2 anaerobic. Aerobic bacteria were present in 86% of the specimens, whereas only anaerobic bacteria were isolated in 7%. The predominant aerobic species were Proteus mirabilis, Staphylococcus haemolyticus and Staphylococcus lugdunensis. The isolated anaerobic bacteria were Dermacoccus nishinomiyaensis and Propionibacterium granulosum. Conclusion A variety of aerobic and anaerobic bacteria was isolated from the HS lesions of our patients. In contrast to previous studies, fewer patients were found to be culture positive, and Staphylococcus aureus was isolated in only 1 of them. More studies are necessary to elucidate the controversial role of bacteria in the pathogenesis of HS. PMID:27170935

  5. Influence of an aniline supplement on the stability of aerobic granular sludge.

    PubMed

    Dai, Yajie; Jiang, Yixin; Su, Haijia

    2015-10-01

    In order to evaluate the stability of aerobic granules in a toxic environment, this study discussed the influence of an aniline supplement on the properties and microbial community of aerobic granules. In the early stages of sequencing batch reactor (SBR) operation, an aniline supplement slightly affected the properties of the aerobic granules (strength, growth rate, SVI and so on). This effect was thereafter removed because of a change in the microbial community and the structure of aerobic granules: with the present of aniline, microbes with biodegradation ability appeared and gathered in the aerobic granules and the aerobic granules densified and settled faster as their SVI decreased to 35 mL/g and settling velocity increased to 41.56 m/h. When a synthetic waste water containing acetate as carbon source was used as influent, aniline (10-500 mg/L) could be degraded in 6 h, at a rate as high as 37.5 mg aniline/(L·h), with a removal rate in excess of 90%, while the effluent COD fell below 100 mg/L from the initial about 2000 mg/L. The aerobic granules cultured by acetate were compact, stable and resistant to aniline. PMID:26233584

  6. Bartonella melophagi in blood of domestic sheep (Ovis aries) and sheep keds (Melophagus ovinus) from the southwestern US: Cultures, genetic characterization, and ecological connections.

    PubMed

    Kosoy, Michael; Bai, Ying; Enscore, Russell; Rizzo, Maria Rosales; Bender, Scott; Popov, Vsevolod; Albayrak, Levent; Fofanov, Yuriy; Chomel, Bruno

    2016-07-15

    Bartonella melophagi sp. nov. was isolated from domestic sheep blood and from sheep keds (Melophagus ovinus) from the southwestern United States. The sequence analyses of the reference strain performed by six molecular markers consistently demonstrated that B. melophagi relates to but differ from other Bartonella species isolated from domestic and wild ruminants. Presence of 183 genes specific for B. melophagi, being absent in genomes of other Bartonella species associated with ruminants also supports the separation of this bacterial species from species of other ruminants. Bartonella DNA was detected in all investigated sheep keds; however, culturing of these bacteria from sheep blood rejects a speculation that B. melophagi is an obligatory endosymbiont. Instead, the results support the hypothesis that the domestic sheep is a natural host reservoir for B. melophagi and the sheep ked its main vector. This bacterium was not isolated from the blood of bighorn sheep and domestic goats belonging to the same subfamily Caprinae. B. melophagi has also been shown to be zoonotic and needs to be investigated further. PMID:27283855

  7. Intrauterine insemination of cultured peripheral blood mononuclear cells prior to embryo transfer improves clinical outcome for patients with repeated implantation failures.

    PubMed

    Madkour, Aicha; Bouamoud, Nouzha; Louanjli, Noureddine; Kaarouch, Ismail; Copin, Henri; Benkhalifa, Moncef; Sefrioui, Omar

    2016-02-01

    Implantation failure is a major limiting factor in assisted reproduction improvement. Dysfunction of embryo-maternal immuno-tolerance pathways may be responsible for repeated implantation failures. This fact is supported by immunotropic theory stipulating that maternal immune cells, essentially uterine CD56+ natural killer cells, are determinants of implantation success. In order to test this hypothesis, we applied endometrium immuno-modulation prior to fresh embryo transfer for patients with repeated implantation failures. Peripheral blood mononuclear cells were isolated from repeated implantation failure patients undergoing assisted reproductive technology cycles. On the day of ovulation induction, cells were isolated and then cultured for 3 days and transferred into the endometrium cavity prior to fresh embryo transfer. This immunotherapy was performed on 27 patients with repeated implantation failures and compared with another 27 patients who served as controls. Implantation and clinical pregnancy were increased significantly in the peripheral blood mononuclear cell test versus control (21.54, 44.44 vs. 8.62, 14.81%). This finding suggests a clear role for endometrium immuno-modulation and the inflammation process in implantation success. Our study showed the feasibility of intrauterine administration of autologous peripheral blood mononuclear cells as an effective therapy to improve clinical outcomes for patients with repeated implantation failures and who are undergoing in vitro fertilization cycles. PMID:25613318

  8. The proliferation potential of promastigotes of the main Leishmania species of the old world in NNN culture medium prepared using blood of four different mammals.

    PubMed

    Ladopoulos, Theodoros; Ntais, Pantelis; Tsirigotakis, Nikolaos; Dokianakis, Emmanouil; Antoniou, Maria

    2015-10-01

    The efficacy of the in vitro cultivation of promastigotes of four Leishmania spp. was tested in the biphasic Novy-MacNeal-Nicolle (NNN) medium prepared using blood from different animals (horse, donkey, goat and sheep). The aim was to test which NNN preparation gave the best yield in the shortest time for different parasite species, in order to obtain a large crop of promastigotes for experimental work and for antigen preparation. Promastigotes of Leishmania infantum, Leishmania donovani, Leishmania tropica and Leishmania major, the four main parasite species occurring in the old world, were defrosted from -80 °C and placed, at equal numbers, in the 4 different NNN preparations. At the end of the 7th day, the NNN medium using horse blood produced the greatest number of promastigotes for all Leishmania spp. tested, whilst goat blood proved the poorest medium, providing culture results only for L. infantum. This finding may be explained by the fact that Leishmania is a nicotinamide adenine dinucleotide (NAD) auxotroph and horse erythrocytes support NAD-dependent microorganisms. PMID:26219203

  9. A facile method to establish human induced pluripotent stem cells from adult blood cells under feeder-free and xeno-free culture conditions: a clinically compliant approach.

    PubMed

    Chou, Bin-Kuan; Gu, Haihui; Gao, Yongxing; Dowey, Sarah N; Wang, Ying; Shi, Jun; Li, Yanxin; Ye, Zhaohui; Cheng, Tao; Cheng, Linzhao

    2015-04-01

    Reprogramming human adult blood mononuclear cells (MNCs) cells by transient plasmid expression is becoming increasingly popular as an attractive method for generating induced pluripotent stem (iPS) cells without the genomic alteration caused by genome-inserting vectors. However, its efficiency is relatively low with adult MNCs compared with cord blood MNCs and other fetal cells and is highly variable among different adult individuals. We report highly efficient iPS cell derivation under clinically compliant conditions via three major improvements. First, we revised a combination of three EBNA1/OriP episomal vectors expressing five transgenes, which increased reprogramming efficiency by ≥10-50-fold from our previous vectors. Second, human recombinant vitronectin proteins were used as cell culture substrates, alleviating the need for feeder cells or animal-sourced proteins. Finally, we eliminated the previously critical step of manually picking individual iPS cell clones by pooling newly emerged iPS cell colonies. Pooled cultures were then purified based on the presence of the TRA-1-60 pluripotency surface antigen, resulting in the ability to rapidly expand iPS cells for subsequent applications. These new improvements permit a consistent and reliable method to generate human iPS cells with minimal clonal variations from blood MNCs, including previously difficult samples such as those from patients with paroxysmal nocturnal hemoglobinuria. In addition, this method of efficiently generating iPS cells under feeder-free and xeno-free conditions allows for the establishment of clinically compliant iPS cell lines for future therapeutic applications. PMID:25742692

  10. Controlled evaluation of BacT/alert standard anaerobic and FAN anaerobic blood culture bottles for the detection of bacteremia and fungemia.

    PubMed Central

    Wilson, M L; Weinstein, M P; Mirrett, S; Reimer, L G; Feldman, R J; Chuard, C R; Reller, L B

    1995-01-01

    FAN medium was formulated to improve microbial recovery, particularly for fastidious microorganisms and for microorganisms causing sepsis in patients receiving antimicrobial therapy. In a controlled clinical evaluation performed at four university-affiliated hospitals, FAN anaerobic bottles were compared with standard anaerobic bottles for yield, speed of detection of microbial growth, and detection of septic episodes. A total of 10,431 blood culture sets were received; both anaerobic bottles of 7,694 blood culture sets were adequately filled with blood. Altogether, 925 isolates were recovered: 557 that were the cause of sepsis, 99 that were indeterminate as the cause of sepsis, and 269 contaminants. More Staphylococcus aureus (P < 0.001), coagulase-negative staphylococci (P < 0.001), Escherichia coli (P < 0.02), and all microorganisms combined (P < 0.005) were recovered from FAN bottles; more nonfermentative gram-negative bacilli (P < 0.05), Torulopsis glabrata (P < 0.001), and other yeasts (P < 0.01) were recovered from standard bottles. Growth of S. aureus (P < 0.001), coagulase-negative staphylococci (P < 0.001), Enterococcus faecalis (P < 0.025), streptococci other than Streptococcus pneumoniae (P < 0.01), and all microorganisms combined (P < 0.001) was detected earlier in standard bottles; growth of more isolates of E. coli (P < 0.05) and anaerobic bacteria (P < 0.01) was detected earlier in FAN bottles. The mean times to detection were 14.2 and 16.1 h for standard and FAN bottles, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7494013

  11. Identification and culture of Kaposi's sarcoma-like spindle cells from the peripheral blood of human immunodeficiency virus-1-infected individuals and normal controls.

    PubMed

    Browning, P J; Sechler, J M; Kaplan, M; Washington, R H; Gendelman, R; Yarchoan, R; Ensoli, B; Gallo, R C

    1994-10-15

    We examined 26 patients with human immunodeficiency virus-1 (HIV-1)-associated Kaposi's sarcoma (KS), and 76 HIV-1-infected (HIV-1+) people without KS or uninfected (HIV-1-) controls for the presence of circulating KS-like spindle cells. Adherent cells that had spindle morphology and several characteristics of spindle cells of KS lesions (KS cells) were identified in the peripheral blood mononuclear cell fraction only after culture in the presence of conditioned medium (CM) from activated lymphocytes. The peripheral blood-derived spindle cells (PBsc) expressed a variety of endothelial cell markers, such as Ulex europaeus I lectin, EN4, EN2/3, EN7/44, CD13, CD34, CD36, CD54, ELAM-1, and HLA-DR. However, they were negative for CD2, CD19, PaIE, and factor VIII-related antigen. The PBsc produced angiogenic factors as evidenced by the ability of CM from these cells to promote growth of normal vascular endothelial cells. In addition, subcutaneously injected PBsc stimulated angiogenesis in vivo in athymic nude mice. We determined that the number of PBsc grown from the peripheral blood of HIV-1+ patients with KS or at high risk to develop KS were increased by 78-fold (P = .0001) and 18-fold (P = .005), respectively, when compared with HIV-1- controls. The number of spindle cells cultured from the HIV-1+ patients at low risk for developing KS, eg, HIV-1+ injection drug users, showed no statistical increase when compared with HIV-1- controls. The presence of increased PBsc with characteristics of KS cells in HIV-1+ KS patients or patients at high risk for developing KS gives insights into the origin of KS cells and may explain the multifocal nature of the disease. In addition, this may be useful in predicting the risk of KS development. PMID:7522639

  12. Soil and sediment bacteria capable of aerobic nitrate respiration.

    PubMed Central

    Carter, J P; Hsaio, Y H; Spiro, S; Richardson, D J

    1995-01-01

    Several laboratory strains of gram-negative bacteria are known to be able to respire nitrate in the presence of oxygen, although the physiological advantage gained from this process is not entirely clear. The contribution that aerobic nitrate respiration makes to the environmental nitrogen cycle has not been studied. As a first step in addressing this question, a strategy which allows for the isolation of organisms capable of reducing nitrate to nitrite following aerobic growth has been developed. Twenty-nine such strains have been isolated from three soils and a freshwater sediment and shown to comprise members of three genera (Pseudomonas, Aeromonas, and Moraxella). All of these strains expressed a nitrate reductase with an active site located in the periplasmic compartment. Twenty-two of the strains showed significant rates of nitrate respiration in the presence of oxygen when assayed with physiological electron donors. Also isolated was one member of the gram-positive genus Arthrobacter, which was likewise able to respire nitrate in the presence of oxygen but appeared to express a different type of nitrate reductase. In the four environments studied, culturable bacteria capable of aerobic nitrate respiration were isolated in significant numbers (10(4) to 10(7) per g of soil or sediment) and in three cases were as abundant as, or more abundant than, culturable bacteria capable of denitrification. Thus, it seems likely that the corespiration of nitrate and oxygen may indeed make a significant contribution to the flux of nitrate to nitrite in the environment. PMID:7487017

  13. Migration, Material Culture, and Identity in William Attaway's "Blood on the Forge" and Harriette Arnow's "The Dollmaker."

    ERIC Educational Resources Information Center

    Morgan, Stacy I.

    2001-01-01

    Discusses how both novels share key thematic elements pertaining to the experiences of migrants from rural Appalachia to multiethnic industrial centers of the urban north. Notes that a focus on the authors' handling of material culture helps to point one with increased clarity and precision to the writerly method by which Attaway and Arnow convey…

  14. "She Has to Drink Blood of the Snake": Culture and Prior Knowledge in Science|Health Education

    ERIC Educational Resources Information Center

    Bricker, Leah A.; Reeve, Suzanne; Bell, Philip

    2014-01-01

    In this analysis, we argue that science education should attend more deeply to youths' cultural resources and practices (e.g. material, social, and intellectual). Inherent in our argument is a call for revisiting conceptions of "prior knowledge" to theorize how people make sense of the complex ecologies of experience, ideas, and…

  15. Selective induction of light chain synthesis in cultures of blood lymphocytes from patients with IgG myelomatosis.

    PubMed Central

    Walker, L A; Johnson, G D; MacLennan, I C

    1988-01-01

    In the present study evidence is provided that neoplastic B cells from the blood from four of 24 patients with myelomatosis were activated selectively with polyclonal B cell mitogens. In three of these patients the activated cells produced light chains without heavy chains; of these, two patients had IgGK paraproteins and one had free lambda light chain disease. The ratio of kappa-expressing to lambda-expressing B cells in the initial blood B cell preparations was within the range for healthy controls for all four patients where neoplastic B cells were selectively activated. It is concluded that in some patients with myelomatosis the neoplastic clone is a mosaic of: (1) cells capable of synthesizing both light and heavy chains with (2) cells producing light chains only. PMID:2832107

  16. Plasma cytokine concentration and the cytokine producing ability of whole blood cell cultures from healthy females with pharmacologically induced hyperprolactinemia.

    PubMed

    Rovenský, J; Lackovic, V; Veselková, Z; Horváthová, M; Koska, J; Blazícková, S; Vigas, M

    1999-01-01

    We investigated the in vitro effect of domperidone-induced hyperprolactinemia on plasma cytokine concentration and blood leukocyte cytokine production in healthy female volunteers. No changes were found in the plasma concentration of interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, interleukin (IL)-4, IL-10, IL-6 and IL-13 during hyperprolactinemia when compared with control values. Using unseparated blood leukocytes, we found that the spontaneous production of IL-6 (4-8 h) and transforming growth factor (TGF)-beta 1 (2-4 h) was significantly decreased and that the in vitro stimulated production of IFN-gamma (2-8 h) and TNF (4 h) was significantly increased compared with control. Our data concerning the increased IFN-gamma and TNF producing capacity of unseparated leukocytes during pharmacologically induced hyperprolactinemia strongly support the possibility that the lymphocyte production of these cytokines can be rapidly amplified by prolactin via a priming mechanism. PMID:10568223

  17. Direct Bacterial Identification in Positive Blood Cultures by Use of Two Commercial Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Systems

    PubMed Central

    Chen, Jonathan H. K.; Ho, Pak-Leung; Kwan, Grace S. W.; She, Kevin K. K.; Siu, Gilman K. H.; Cheng, Vincent C. C.; Yuen, Kwok-Yung

    2013-01-01

    Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) for the identification of bacteria and fungi was recently introduced in microbiology laboratories. This technology could greatly improve the clinical management of patients and guidance for chemotherapy. In this study, we used a commercial MALDI Sepsityper extraction method to evaluate the performance of two commercial MALDI-TOF MS systems, the Vitek MS IVD (bioMérieux) and the Microflex LT Biotyper (Bruker Daltonics) for direct bacterial identification in positive blood cultures. In 181 monomicrobial cultures, both systems generated genus to species level identifications for >90% of the specimens (Biotyper, 177/181 [97.8%]; Vitek MS IVD, 167/181 [92.3%]). Overall, the Biotyper system generated significantly more accurate identifications than the Vitek MS IVD system (P = 0.016; 177 versus 167 out of 181 specimens). The Biotyper system identified the minority species among polymicrobial blood cultures. We also compared the performance of an in-house extraction method with that of the Sepsityper on both MALDI-TOF MS systems. The in-house method generated more correct identifications at the genus level than the Sepsityper (96.7% versus 93.5%) on the Biotyper system, whereas the two methods exhibited the same performance level (88.0% versus 88.0%) on the Vitek MS IVD system. Our study confirmed the practical advantages of MALDI-TOF MS, and our in-house extraction method reduced the reagent cost to $1 per specimen, with a shorter turnaround time of 3 h, which is highly cost-effective for a diagnostic microbiology service. PMID:23515548

  18. Association of body mass index and aerobic physical fitness with cardiovascular risk factors in children☆

    PubMed Central

    Gonçalves, Reginaldo; Szmuchrowski, Leszek Antony; Damasceno, Vinícius Oliveira; de Medeiros, Marcelo Lemos; Couto, Bruno Pena; Lamounier, Joel Alves

    2014-01-01

    Objective: To identify the association between both, body mass index and aerobic fitness, with cardiovascular disease risk factors in children. Methods: Cross-sectional study, carried out in Itaúna-MG, in 2010, with 290 school children ranging from 6 to 10 years-old of both sexes, randomly selected. Children from schools located in the countryside and those with medical restrctions for physical activity were not included. Blood sample was collected after a 12-hour fasting period. Blood pressure, stature and weight were evaluated in accordance with international standards. The following were considered as cardiovascular risk factors: high blood pressure, high total cholesterol, LDL, triglycerides and insulin levels, and low HDL. The statistical analysis included the Spearman's coefficient and the logistic regression, with cardiovascular risk factors as dependent variables. Results: Significant correlations were found, in both sexes, among body mass index and aerobic fitness with most of the cardiovascular risk factors. Children of both sexes with body mass index in the fourth quartile demonstrated increased chances of having high blood insulin and clustering cardiovascular risk factors. Moreover, girls with aerobic fitness in the first quartile also demonstrated increased chances of having high blood insulin and clustering cardiovascular risk factors. Conclusion: The significant associations and the increased chances of having cardiovascular risk factors in children with less aerobic fitness and higher levels of body mass index justify the use of these variables for health monitoring in Pediatrics. PMID:25479851

  19. Identifying viable regulatory and innovation pathways for regenerative medicine: a case study of cultured red blood cells.

    PubMed

    Mittra, J; Tait, J; Mastroeni, M; Turner, M L; Mountford, J C; Bruce, K

    2015-01-25

    The creation of red blood cells for the blood transfusion markets represents a highly innovative application of regenerative medicine with a medium term (5-10 year) prospect for first clinical studies. This article describes a case study analysis of a project to derive red blood cells from human embryonic stem cells, including the systemic challenges arising from (i) the selection of appropriate and viable regulatory protocols and (ii) technological constraints related to stem cell manufacture and scale up to clinical Good Manufacturing Practice (GMP) standard. The method used for case study analysis (Analysis of Life Science Innovation Systems (ALSIS)) is also innovative, demonstrating a new approach to social and natural science collaboration to foresight product development pathways. Issues arising along the development pathway include cell manufacture and scale-up challenges, affected by regulatory demands emerging from the innovation ecosystem (preclinical testing and clinical trials). Our discussion reflects on the efforts being made by regulators to adapt the current pharmaceuticals-based regulatory model to an allogeneic regenerative medicine product and the broader lessons from this case study for successful innovation and translation of regenerative medicine therapies, including the role of methodological and regulatory innovation in future development in the field. PMID:25094050

  20. Chemical characterization of fatty acids, alkanes, n-diols and alkyl esters produced by a mixed culture of Trichoderma koningii and Penicillium janthinellum grown aerobically on undecanoic acid, potatoe dextrose and their mixture.

    PubMed

    Monreal, Carlos M; Chahal, Amarpreet; Schnitzer, Morris; Rowland, Owen

    2016-01-01

    Little is known about the mixed fungal synthesis of high-value aliphatics derived from the metabolism of simple and complex carbon substrates. Trichoderma koningii and Penicillium janthinellum were fed with undecanoic acid (UDA), potatoe dextrose broth (PDB), and their mixture. Pyrolysis Field Ionization Mass Spectrometry (Py-FIMS) together with (1)H and (13)C Nuclear Magnetic Resonance (NMR) characterized CHCl3 soluble aliphatics in the fungal cell culture. Data from NMR and Py-FIMS analysis were complementary to each other. On average, the mixed fungal species produced mostly fatty acids (28% of total ion intensity, TII) > alkanes (2% of TII) > n-diols (2% of TII) > and alkyl esters (0.8% of TII) when fed with UDA, PDB or UDA+PDB. The cell culture accumulated aliphatics extracellularly, although most of the identified compounds accumulated intracellularly. The mixed fungal culture produced high-value chemicals from the metabolic conversion of simple and complex carbon substrates. PMID:26852878

  1. In Vitro Differentiation of Human Umbilical Cord Blood CD133+Cells into Insulin Producing Cells in Co-Culture with Rat Pancreatic Mesenchymal Stem Cells

    PubMed Central

    Sahraneshin Samani, Fazel; Ebrahimi, Marzieh; Zandieh, Tahereh; Khoshchehreh, Reyhaneh; Baghaban Eslaminejad, Mohamadreza; Aghdami, Nasser; Baharvand, Hossein

    2015-01-01

    Objective Pancreatic stroma plays an important role in the induction of pancreatic cells by the use of close range signaling. In this respect, we presume that pancreatic mesenchymal cells (PMCs) as a fundamental factor of the stromal niche may have an effective role in differentiation of umbilical cord blood cluster of differentiation 133+ (UCB-CD133+) cells into newly-formed β-cells in vitro. Materials and Methods This study is an experimental research. The UCB-CD133+cells were purified by magnetic activated cell sorting (MACS) and differentiated into insulin producing cells (IPCs) in co-culture, both directly and indirectly with rat PMCs. Immunocytochemistry and enzyme linked immune sorbent assay (ELISA) were used to determine expression and production of insulin and C-peptide at the protein level. Results Our results demonstrated that UCB-CD133+differentiated into IPCs. Cells in islet-like clusters with (out) co-cultured with rat pancreatic stromal cells produced insulin and C-peptide and released them into the culture medium at the end of the induction protocol. However they did not respond well to glucose challenges. Conclusion Rat PMCs possibly affect differentiation of UCB-CD133+cells into IPCs by increasing the number of immature β-cells. PMID:26199900

  2. `She Has to Drink Blood of the Snake': Culture and prior knowledge in science|health education

    NASA Astrophysics Data System (ADS)

    Bricker, Leah A.; Reeve, Suzanne; Bell, Philip

    2014-06-01

    In this analysis, we argue that science education should attend more deeply to youths' cultural resources and practices (e.g. material, social, and intellectual). Inherent in our argument is a call for revisiting conceptions of 'prior knowledge' to theorize how people make sense of the complex ecologies of experience, ideas, and cultural practices that undergird any learning moment. We illustrate our argument using examples from the domain of personal health, chosen because of its tremendous societal impact and its significant areas of overlap with biology, chemistry, physics, and other scientific disciplines taught as core subjects in schools. Using data from a team ethnography of young people's science and technology learning across settings and over developmental timescales, we highlight two youths' experiences and understandings related to personal health, and how those experiences and understandings influenced the youths' sense-making about the natural world. We then discuss the implications of our argument for science education.

  3. Aerobic granular processes: Current research trends.

    PubMed

    Zhang, Quanguo; Hu, Jianjun; Lee, Duu-Jong

    2016-06-01

    Aerobic granules are large biological aggregates with compact interiors that can be used in efficient wastewater treatment. This mini-review presents new researches on the development of aerobic granular processes, extended treatments for complicated pollutants, granulation mechanisms and enhancements of granule stability in long-term operation or storage, and the reuse of waste biomass as renewable resources. A discussion on the challenges of, and prospects for, the commercialization of aerobic granular process is provided. PMID:26873285

  4. Stable carbon isotope fractionation during aerobic biodegradation of chlorinated ethenes

    SciTech Connect

    Chu, Kung-Hui; Mahendra, Shaily; Song, Donald L.; Conrad, Mark E.; Alvarez-Cohen, Lisa

    2003-06-01

    Stable isotope analysis is recognized as a powerful tool for monitoring, assessing, and validating in-situ bioremediation processes. In this study, kinetic carbon isotope fractionation factors () associated with the aerobic biodegradation of vinyl chloride (VC), cis-1,2-dichloroethylene (cDCE), and trichloroethylene (TCE) were examined. Of the three solvents, the largest fractionation effects were observed for biodegradation of VC. Both metabolic and cometabolic VC degradation were studied using Mycobacterium aurum L1 (grown on VC), Methylosinus trichosporium OB3b (grown on methane), Mycobacterium vaccae JOB 5 (grown on propane), and two VC enrichment cultures seeded from contaminated soils of Alameda Point and Travis Air Force Base, CA. M. aurum L1 caused the greatest fractionation (= -5.7) while for the cometabolic cultures, values ranged from -3.2 to -4.8. VC fractionation patterns for the enrichment cultures were within the range of those observed for the metabolic and cometabolic cultures (= -4.5 to -5.5). The fractionation for cometabolic degradation of TCE by Me. trichosporium OB3b was low (= -1.1), while no quantifiable carbon isotopic fractionation was observed during the cometabolic degradation of cDCE. For all three of the tested chlorinated ethenes, isotopic fractionation measured during aerobic degradation was significantly smaller than that reported for anaerobic reductive dechlorination. This study suggests that analysis of compound-specific isotopic fractionation could assist in determining whether aerobic or anaerobic degradation of VC and cDCE predominates in field applications of in-situ bioremediation. In contrast, isotopic fractionation effects associated with metabolic and cometabolic reactions are not sufficiently dissimilar to distinguish these processes in the field.

  5. A Subset of Circulating Blood Mycobacteria-Specific CD4 T Cells Can Predict the Time to Mycobacterium tuberculosis Sputum Culture Conversion

    PubMed Central

    Lugongolo, Masixole; Gwala, Thabisile; Kiravu, Agano; Deniso, Pamela; Stewart-Isherwood, Lynsey; Omar, Shaheed Vally; Grobusch, Martin P.; Coetzee, Gerrit; Conradie, Francesca; Ismail, Nazir; Kaplan, Gilla; Fallows, Dorothy

    2014-01-01

    We investigated 18 HIV-negative patients with MDR-TB for M. tuberculosis (Mtb)- and PPD-specific CD4 T cell responses and followed them over 6 months of drug therapy. Twelve of these patients were sputum culture (SC) positive and six patients were SC negative upon enrollment. Our aim was to identify a subset of mycobacteria-specific CD4 T cells that would predict time to culture conversion. The total frequency of mycobacteria-specific CD4 T cells at baseline could not distinguish patients showing positive or negative SC. However, a greater proportion of late-differentiated (LD) Mtb- and PPD-specific memory CD4 T cells was found in SC positive patients than in those who were SC negative (p = 0.004 and p = 0.0012, respectively). Similarly, a higher co-expression of HLA-DR+Ki67+ on Mtb- and PPD-specific CD4 T cells could also discriminate between sputum SC positive versus SC negative (p = 0.004 and p = 0.001, respectively). Receiver operating characteristic (ROC) analysis revealed that baseline levels of Ki67+HLA-DR+ Mtb- and PPD-specific CD4 T cells were predictive of the time to sputum culture conversion, with area-under-the-curve of 0.8 (p = 0.027). Upon treatment, there was a significant decline of these Ki67+HLA-DR+ T cell populations in the first 2 months, with a progressive increase in mycobacteria-specific polyfunctional IFNγ+IL2+TNFα+ CD4 T cells over 6 months. Thus, a subset of activated and proliferating mycobacterial-specific CD4 T cells (Ki67+HLA-DR+) may provide a valuable marker in peripheral blood that predicts time to sputum culture conversion in TB patients at the start of treatment. PMID:25048802

  6. A subset of circulating blood mycobacteria-specific CD4 T cells can predict the time to Mycobacterium tuberculosis sputum culture conversion.

    PubMed

    Riou, Catherine; Gray, Clive M; Lugongolo, Masixole; Gwala, Thabisile; Kiravu, Agano; Deniso, Pamela; Stewart-Isherwood, Lynsey; Omar, Shaheed Vally; Grobusch, Martin P; Coetzee, Gerrit; Conradie, Francesca; Ismail, Nazir; Kaplan, Gilla; Fallows, Dorothy

    2014-01-01

    We investigated 18 HIV-negative patients with MDR-TB for M. tuberculosis (Mtb)- and PPD-specific CD4 T cell responses and followed them over 6 months of drug therapy. Twelve of these patients were sputum culture (SC) positive and six patients were SC negative upon enrollment. Our aim was to identify a subset of mycobacteria-specific CD4 T cells that would predict time to culture conversion. The total frequency of mycobacteria-specific CD4 T cells at baseline could not distinguish patients showing positive or negative SC. However, a greater proportion of late-differentiated (LD) Mtb- and PPD-specific memory CD4 T cells was found in SC positive patients than in those who were SC negative (p = 0.004 and p = 0.0012, respectively). Similarly, a higher co-expression of HLA-DR+ Ki67+ on Mtb- and PPD-specific CD4 T cells could also discriminate between sputum SC positive versus SC negative (p = 0.004 and p = 0.001, respectively). Receiver operating characteristic (ROC) analysis revealed that baseline levels of Ki67+ HLA-DR+ Mtb- and PPD-specific CD4 T cells were predictive of the time to sputum culture conversion, with area-under-the-curve of 0.8 (p = 0.027). Upon treatment, there was a significant decline of these Ki67+ HLA-DR+ T cell populations in the first 2 months, with a progressive increase in mycobacteria-specific polyfunctional IFNγ+ IL2+ TNFα+ CD4 T cells over 6 months. Thus, a subset of activated and proliferating mycobacterial-specific CD4 T cells (Ki67+ HLA-DR+) may provide a valuable marker in peripheral blood that predicts time to sputum culture conversion in TB patients at the start of treatment. PMID:25048802

  7. Optimized Use of the MALDI BioTyper System and the FilmArray BCID Panel for Direct Identification of Microbial Pathogens from Positive Blood Cultures

    PubMed Central

    Fiori, B.; D'Inzeo, T.; Giaquinto, A.; Menchinelli, G.; Liotti, F. M.; de Maio, F.; De Angelis, G.; Quaranta, G.; Nagel, D.; Tumbarello, M.; Sanguinetti, M.

    2015-01-01

    Despite the current reliance on blood cultures (BCs), the diagnosis of bloodstream infections (BSIs) can be sped up using new technologies performed directly on positive BC bottles. Two methods (the MALDI BioTyper system and FilmArray blood culture identification [BCID] panel) are potentially applicable. In this study, we performed a large-scale clinical evaluation (1,585 microorganisms from 1,394 BSI episodes) on the combined use of the MALDI BioTyper and FilmArray BCID panel compared to a reference (culture-based) method. As a result, the causative organisms of 97.7% (1,362/1,394) of the BSIs were correctly identified by our MALDI BioTyper and FilmArray BCID-based algorithm. Specifically, 65 (5.3%) out of 1,223 monomicrobial BCs that provided incorrect or invalid identifications with the MALDI BioTyper were accurately detected by the FilmArray BCID panel; additionally, 153 (89.5%) out of 171 polymicrobial BCs achieved complete identification with the FilmArray BCID panel. Conversely, full use of the MALDI BioTyper would have resulted in the identification of only 1 causative organism in 97/171 (56.7%) of the polymicrobial cultures. By applying our diagnostic algorithm, the median time to identification was shortened (19.5 h versus 41.7 h with the reference method; P < 0.001), and the minimized use of the FilmArray BCID panel led to a significant cost savings. Twenty-six out of 31 microorganisms that could not be identified were species/genera not designed to be detected with the FilmArray BCID panel, indicating that subculture was not dispensable for a few of our BSI episodes. In summary, the fast and effective testing of BC bottles is realistically adoptable in the clinical microbiology laboratory workflow, although the usefulness of this testing for the management of BSIs remains to be established. PMID:26677254

  8. Use of PNA FISH for blood cultures growing Gram-positive cocci in chains without a concomitant antibiotic stewardship intervention does not improve time to appropriate antibiotic therapy.

    PubMed

    Cosgrove, Sara E; Li, David X; Tamma, Pranita D; Avdic, Edina; Hadhazy, Eric; Wakefield, Teresa; Gherna, Michael; Carroll, Karen C

    2016-09-01

    Peptide nucleic acid fluorescence in situ hybridization (PNA FISH) is a rapid diagnostic assay that can identify certain organisms growing in blood cultures 30-90 min from the time of positive Gram-stain. Existing studies have demonstrated a clinical utility with this assay when antibiotic stewardship programs assist clinicians with interpreting the results. However, the benefit of these rapid assays in the absence of concomitant antibiotic stewardship involvement is unclear. In this randomized study of 220 patients with enterococcal or streptococcal bacteremia, we found that PNA FISH, in the absence of concomitant input from an antibiotic stewardship program, had no impact on time to effective or optimal therapy, length of hospital stay, or in-hospital mortality. Our results suggest that in the absence of guidance from an antibiotic stewardship program, the clinical benefits of rapid diagnostic microbiological tools may be reduced. PMID:27412814

  9. Antibiotic treatment algorithm development based on a microarray nucleic acid assay for rapid bacterial identification and resistance determination from positive blood cultures.

    PubMed

    Rödel, Jürgen; Karrasch, Matthias; Edel, Birgit; Stoll, Sylvia; Bohnert, Jürgen; Löffler, Bettina; Saupe, Angela; Pfister, Wolfgang

    2016-03-01

    Rapid diagnosis of bloodstream infections remains a challenge for the early targeting of an antibiotic therapy in sepsis patients. In recent studies, the reliability of the Nanosphere Verigene Gram-positive and Gram-negative blood culture (BC-GP and BC-GN) assays for the rapid identification of bacteria and resistance genes directly from positive BCs has been demonstrated. In this work, we have developed a model to define treatment recommendations by combining Verigene test results with knowledge on local antibiotic resistance patterns of bacterial pathogens. The data of 275 positive BCs were analyzed. Two hundred sixty-three isolates (95.6%) were included in the Verigene assay panels, and 257 isolates (93.5%) were correctly identified. The agreement of the detection of resistance genes with subsequent phenotypic susceptibility testing was 100%. The hospital antibiogram was used to develop a treatment algorithm on the basis of Verigene results that may contribute to a faster patient management. PMID:26712265

  10. [Evaluation of peptide nucleic acid fluorescent in situ hybridization (PNA FISH) method in the identifi cation of Candida species isolated from blood cultures].

    PubMed

    Aydemir, Gonca; Koç, Ayşe Nedret; Atalay, Mustafa Altay

    2016-04-01

    In recent years, increased number of patients who are hospitalized in intensive care units, received immunosuppressive therapy and treated with broad-spectrum antibiotics that can lead an increase in the incidence of systemic candidiasis. In these patients, the most common clinical manifestation is candidemia. Since the identification of Candida species isolated from blood cultures is time consuming by conventional (morphological and biochemical) methods, rapid, reliable and accurate methods are needed. For this purpose novel systems have been developed to identify the agent directly. The aim of this study was to evaluate the peptide nucleic acid fluorescent in situ hybridization (PNA FISH) method for the identification of Candida species by comparing with the conventional methods. A total of 50 patients who were admitted to Erciyes University Medical Faculty Hospital clinics and followed with prediagnosis of systemic fungal infections whose blood cultures were positive for the yeasts between July 2011 and July 2012 were included in the study. The conventional identification of Candida isolates was performed by considering macroscopic and microscopic morphology, germ tube test, cycloheximide sensitivity, urease activity and carbohydrate assimilation patterns with API 20C AUX (bioMerieux, France) test. PNA FISH method was conducted by the use of a commercial kit namely Yeast Traffic Light(®) PNA FISH (AdvanDx, USA). According to morphological and biochemical characteristics (conventional methods), 19 (38%) out of 50 Candida isolates were identified as C.albicans, 12 (24%) as C.glabrata, five (10%) as C.parapsilosis, five (10%) as C.kefyr, four (8%) as C.krusei, two (4%) as C.guilliermondii, two (4%) as C.tropicalis and one (2%) as C.lusitaniae. On the other hand, 24 (48%) of the isolates were identified as C.albicans/C.parapsilosis (with green fluorescence), 16 (32%) as C.glabrata/C.krusei (with red fluorescence) and one (%2) as C.tropicalis (with yellow

  11. High-Yield Method for Isolation and Culture of Endothelial Cells from Rat Coronary Blood Vessels Suitable for Analysis of Intracellular Calcium and Nitric Oxide Biosynthetic Pathways

    PubMed Central

    Nistri, Silvia; Mazzetti, Luca; Failli, Paola

    2002-01-01

    We describe here a method for isolating endothelial cells from rat heart blood vessels by means of coronary microperfusion with collagenase. This methods makes it possible to obtain high amounts of endothelial cells in culture which retain the functional properties of their in vivo counterparts, including the ability to uptake fluorescently-labeled acetylated low-density lipoproteins and to respond to vasoactive agents by modulating intracellular calcium and by upregulating intrinsic nitric oxide generation. The main advantages of our technique are: (i) good reproducibility, (ii) accurate sterility that can be maintained throughout the isolation procedure and (iii) high yield of pure endothelial cells, mainly due to microperfusion and temperature-controlled incubation with collagenase which allow an optimal distribution of this enzyme within the coronary vascular bed. PMID:12734571

  12. Maximal aerobic exercise following prolonged sleep deprivation.

    PubMed

    Goodman, J; Radomski, M; Hart, L; Plyley, M; Shephard, R J

    1989-12-01

    The effect of 60 h without sleep upon maximal oxygen intake was examined in 12 young women, using a cycle ergometer protocol. The arousal of the subjects was maintained by requiring the performance of a sequence of cognitive tasks throughout the experimental period. Well-defined oxygen intake plateaus were obtained both before and after sleep deprivation, and no change of maximal oxygen intake was observed immediately following sleep deprivation. The endurance time for exhausting exercise also remained unchanged, as did such markers of aerobic performance as peak exercise ventilation, peak heart rate, peak respiratory gas exchange ratio, and peak blood lactate. However, as in an earlier study of sleep deprivation with male subjects (in which a decrease of treadmill maximal oxygen intake was observed), the formula of Dill and Costill (4) indicated the development of a substantial (11.6%) increase of estimated plasma volume percentage with corresponding decreases in hematocrit and red cell count. Possible factors sustaining maximal oxygen intake under the conditions of the present experiment include (1) maintained arousal of the subjects with no decrease in peak exercise ventilation or the related respiratory work and (2) use of a cycle ergometer rather than a treadmill test with possible concurrent differences in the impact of hematocrit levels and plasma volume expansion upon peak cardiac output and thus oxygen delivery to the working muscles. PMID:2628360

  13. Aerobic fitness and orthostatic tolerance: Evidence against an association

    NASA Technical Reports Server (NTRS)

    Ebert, Thomas J.

    1994-01-01

    This presentation will focus on only one side of the debate as to whether high levels of aerobic fitness have a deleterious effect on tolerance to gravitational stress. This issue was raised in the early 1970's as a result of two research publications. The first work investigated the carotid sinus baroreflex of humans with an airtight chamber that surrounded the head and neck. The steady-state reflex changes in blood pressure that were recorded 3 minutes after application of the head and neck stimuli, were attenuated in an athletic group compared to a sedentary group of volunteers. A second report in the NASA literature indicated that five endurance-trained runners were less tolerant to LBNP than five nonrunners. These early research findings have stimulated a considerable amount of interest that has lead to a growing number of research efforts seeking an association between aerobic fitness and orthostatic tolerance in humans. I will briefly review some of the more pertinent published research information which suggests that there is no relationship between aerobic fitness and orthostatic tolerance in humans.

  14. Preliminary identification of beta-hemolytic streptococci in throat swab cultures with a commercial blood agar slide (streptocult).

    PubMed Central

    Christensen, P; Danielsson, D; Hovelius, B; Kjellander, J

    1982-01-01

    A commercial blood agar slide (Streptocult, Orion Diagnostica) was used for preliminary identification of beta-hemolytic streptococci of groups A, C, and G in throat swab specimens. The sensitivity of the test was 93.6% and the specificity was 94.7%, as judged from 580 specimens. A model is suggested for routine processing of throat swab specimens, involving inoculation and reading of the slide in general practice and transport of positive or inconclusive slides to a bacteriology laboratory for isolation and serological grouping of beta-hemolytic streptococci. The model combines preliminary detection of beta-hemolytic streptococci within 24 h with the reliability of serological groupings, and should reduce the volume of specimens sent to the laboratory considerably. Images PMID:7050155

  15. Staph ID/R: a Rapid Method for Determining Staphylococcus Species Identity and Detecting the mecA Gene Directly from Positive Blood Culture

    PubMed Central

    Pasko, Chris; Dunn, John; Jaeckel, Heidi; Nieuwlandt, Dan; Weed, Diane; Woodruff, Evelyn; Zheng, Xiaotian

    2012-01-01

    Rapid diagnosis of staphylococcal bacteremia directs appropriate antimicrobial therapy, leading to improved patient outcome. We describe herein a rapid test (<75 min) that can identify the major pathogenic strains of Staphylococcus to the species level as well as the presence or absence of the methicillin resistance determinant gene, mecA. The test, Staph ID/R, combines a rapid isothermal nucleic acid amplification method, helicase-dependent amplification (HDA), with a chip-based array that produces unambiguous visible results. The analytic sensitivity was 1 CFU per reaction for the mecA gene and was 1 to 250 CFU per reaction depending on the staphylococcal species present in the positive blood culture. Staph ID/R has excellent specificity as well, with no cross-reactivity observed. We validated the performance of Staph ID/R by testing 104 frozen clinical positive blood cultures and comparing the results with rpoB gene or 16S rRNA gene sequencing for species identity determinations and mecA gene PCR to confirm mecA gene results. Staph ID/R agreed with mecA gene PCR for all samples and agreed with rpoB/16S rRNA gene sequencing in all cases except for one sample that contained a mixture of two staphylococcal species, one of which Staph ID/R correctly identified, for an overall agreement of 99.0% (P < 0.01). Staph ID/R could potentially be used to positively affect patient management for Staphylococcus-mediated bacteremia. PMID:22170912

  16. Evaluation of PCR-Reverse Blot Hybridization Assay, REBA Sepsis-ID Test, for Simultaneous Identification of Bacterial Pathogens and mecA and van Genes from Blood Culture Bottles

    PubMed Central

    Park, Soon Deok; Lee, Gyusang; Wang, Hye-young; Park, Min; Kim, Sunghyun; Kim, Hyunjung; Kim, Jungho; Kim, Young Keun; Kim, Hyo Youl; Lee, Hyeyoung

    2014-01-01

    Background The aim of this study was to evaluate a newly developed PCR-based reverse blot hybridization assay (PCR-REBA), REBA Sepsis-ID (M&D, Wonju, Korea), to rapidly detect the presence of bacteremia and antimicrobial resistance gene in blood culture samples. Methods One thousand four hundred consecutive blood culture samples from patients with a delta neutrophil index greater than 2.7% were selected from March to July in 2013. Three hundred positive and 1,100 negative for bacterial growth in blood culture bottles samples were tested by conventional and real-time PCR-REBA, respectively. Results The overall agreement between the conventional identification test and the REBA Sepsis-ID test was 95.3% (286/300). Agreement for gram-positive bacteria, gram-negative bacteria, fungi, and polymicrobials was 94.5% (190/201), 97.3% (71/73), 100% (14/14), and 91.7% (11/12), respectively. The detection rate of the mecA gene from methicillin-resistant Staphylococcus isolates was 97.8% (90/92). The vanA gene was detected in one blood culture sample from which vancomycin-resistant Enterococcus was isolated. When the cycle threshold for real-time PCR was defined as 30.0, 2.4% (26/1,100) of negative blood culture samples tested positive by real-time PCR. Conclusions The REBA Sepsis-ID test is capable of simultaneously and quickly detecting both causative agents and antimicrobial resistance genes, such as mecA and van, in blood culture positive samples. PMID:25368820

  17. Aerobic fitness ecological validity in elite soccer players: a metabolic power approach.

    PubMed

    Manzi, Vincenzo; Impellizzeri, Franco; Castagna, Carlo

    2014-04-01

    The aim of this study was to examine the association between match metabolic power (MP) categories and aerobic fitness in elite-level male soccer players. Seventeen male professional soccer players were tested for VO2max, maximal aerobic speed (MAS), VO2 at ventilatory threshold (VO2VT and %VO2VT), and speed at a selected blood lactate concentration (4 mmol·L(-1), V(L4)). Aerobic fitness tests were performed at the end of preseason and after 12 and 24 weeks during the championship. Aerobic fitness and MP variables were considered as mean of all seasonal testing and of 16 Championship home matches for all the calculations, respectively. Results showed that VO2max (from 0.55 to 0.68), MAS (from 0.52 to 0.72), VO2VT (from 0.72 to 0.83), %VO2maxVT (from 0.62 to 0.65), and V(L4) (from 0.56 to 0.73) were significantly (p < 0.05 to 0.001) large to very large associated with MP variables. These results provide evidence to the ecological validity of aerobic fitness in male professional soccer. Strength and conditioning professionals should consider aerobic fitness in their training program when dealing with professional male soccer players. The MP method resulted an interesting approach for tracking external load in male professional soccer players. PMID:24345968

  18. Specific Accumulation of Tumor-Derived Adhesion Factor in Tumor Blood Vessels and in Capillary Tube-Like Structures of Cultured Vascular Endothelial Cells

    NASA Astrophysics Data System (ADS)

    Akaogi, Kotaro; Okabe, Yukie; Sato, Junji; Nagashima, Yoji; Yasumitsu, Hidetaro; Sugahara, Kazuyuki; Miyazaki, Kaoru

    1996-08-01

    Tumor-derived adhesion factor (TAF) was previously identified as a cell adhesion molecule secreted by human bladder carcinoma cell line EJ-1. To elucidate the physiological function of TAF, we examined its distribution in human normal and tumor tissues. Immunochemical staining with an anti-TAF monoclonal antibody showed that TAF was specifically accumulated in small blood vessels and capillaries within and adjacent to tumor nests, but not in those in normal tissues. Tumor blood vessel-specific staining of TAF was observed in various human cancers, such as esophagus, brain, lung, and stomach cancers. Double immunofluorescent staining showed apparent colocalization of TAF and type IV collagen in the vascular basement membrane. In vitro experiments demonstrated that TAF preferentially bound to type IV collagen among various extracellular matrix components tested. In cell culture experiments, TAF promoted adhesion of human umbilical vein endothelial cells to type IV collagen substrate and induced their morphological change. Furthermore, when the endothelial cells were induced to form capillary tube-like structures by type I collagen, TAF and type IV collagen were exclusively detected on the tubular structures. The capillary tube formation in vitro was prevented by heparin, which inhibited the binding of TAF to the endothelial cells. These results strongly suggest that TAF contributes to the organization of new capillary vessels in tumor tissues by modulating the interaction of endothelial cells with type IV collagen.

  19. Comparison of active dry yeast (Saccharomyces cerevisiae) and yeast culture for growth performance, carcass traits, meat quality and blood indexes in finishing bulls.

    PubMed

    Geng, Chun-Yin; Ren, Li-Ping; Zhou, Zhen-Ming; Chang, Ying; Meng, Qing-Xiang

    2016-08-01

    This study was conducted to compare the effect of active dry yeasts (ADY) and yeast cultures (YC), two typical products of yeast preparations, on growth performance, carcass characteristics, meat quality and blood indexes in finishing bulls fed a high-concentrate diet. Forty-five finishing bulls (mean body weight (BW) ± standard deviation: 505 ± 29 kg BW) were allocated to three groups of 15 bulls and assigned randomly to one of three diets which were CON diet (basal diet), ADY diet (basal diet + Levucell SC) and YC diet (basal diet + Diamond V XP), respectively. After 98 days of trial, all bulls were slaughtered. The result showed that ADY rather than YC improved growth performance and carcass traits of bulls compared to CON. Moreover, both ADY and YC improved beef tenderness and changed blood indexes related to fat metabolism. In conclusion, ADY had more pronounced effect on growth performance of bulls fed high-concentrate diet, and both ADY and YC improved the beef quality by intensive fat metabolism. PMID:26472702

  20. Influence of Hydration Status on Changes in Plasma Cortisol, Leukocytes, and Antigen-Stimulated Cytokine Production by Whole Blood Culture following Prolonged Exercise.

    PubMed

    Svendsen, Ida S; Killer, Sophie C; Gleeson, Michael

    2014-01-01

    Elevated antigen-stimulated anti-inflammatory cytokine production appears to be a risk factor for upper respiratory tract illness in athletes. The purpose of this study was to determine the effects of prolonged exercise and hydration on antigen-stimulated cytokine production. Twelve healthy males cycled for 120 min at 60% [Formula: see text] on two occasions, either euhydrated or moderately hypohydrated (induced by fluid restriction for 24 h). Blood samples were collected before and after exercise and following 2 h recovery for determination of cell counts, plasma cortisol, and in vitro antigen-stimulated cytokine production by whole blood culture. Fluid restriction resulted in mean body mass loss of 1.3% and 3.9% before and after exercise, respectively. Exercise elicited a significant leukocytosis and elevated plasma cortisol, with no differences between trials. IL-6 production was significantly reduced 2 h postexercise (P < 0.05), while IL-10 production was elevated postexercise (P < 0.05). IFN- γ and IL-2 production tended to decrease postexercise. No significant effect of hydration status was observed for the measured variables. Prolonged exercise appears to result in augmented anti-inflammatory cytokine release in response to antigen challenge, possibly coupled with acute suppression of proinflammatory cytokine production, corresponding with studies using mitogen or endotoxin as stimulant. Moderate hypohydration does not appear to influence these changes. PMID:24967270

  1. Direct identification of microorganisms from positive blood cultures using the lysis-filtration technique and matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS): a multicentre study.

    PubMed

    Farina, Claudio; Arena, Fabio; Casprini, Patrizia; Cichero, Paola; Clementi, Massimo; Cosentino, Marina; Degl'Innocenti, Roberto; Giani, Tommaso; Luzzaro, Francesco; Mattei, Romano; Mauri, Carola; Nardone, Maria; Rossolini, Gian Maria; Serna Ortega, Paula Andrea; Vailati, Francesca

    2015-04-01

    Microbial identification from blood cultures is essential to institute optimal antibiotic therapy and improve survival possibilities. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been successfully applied to identify bacteria and yeasts from positive blood cultures broths. The aim of this multicentre study was to evaluate the reliability of the lysis-filtration technique associated with MALDI-TOF MS to directly identify microorganisms from 765 positive blood cultures collected in six Italian hospitals. Overall, 675/765 (78.1%) blood isolates were correctly identified at the species level, with significant differences between Gram-negative and Gram-positive bacteria (92.6%, and 69.8%, respectively). Some difficulties arise in identifying Streptococcus pneumoniae, Staphylococcus aureus, yeasts and anaerobes. The lysis-filtration protocol is a suitable procedure in terms of performance in identifying microorganisms, but it is quite expensive and technically time-consuming since the time of filtration is not regular for all the samples. The application of the MALDI-TOF MS technique to the direct microbial identification from positive blood cultures is a very promising approach, even if more experience must be gained to minimize errors and costs. PMID:25938749

  2. Cardiovascular and behavioral effects of aerobic exercise training in healthy older men and women.

    PubMed

    Blumenthal, J A; Emery, C F; Madden, D J; George, L K; Coleman, R E; Riddle, M W; McKee, D C; Reasoner, J; Williams, R S

    1989-09-01

    The cardiovascular and behavioral adaptations associated with a 4-month program of aerobic exercise training were examined in 101 older men and women (mean age = 67 years). Subjects were randomly assigned to an Aerobic Exercise group, a Yoga and Flexibility control group, or a Waiting List control group. Prior to and following the 4-month program, subjects underwent comprehensive physiological and psychological evaluations. Physiological measures included measurement of blood pressure, lipids, bone density, and cardiorespiratory fitness including direct measurements of peak oxygen consumption (VO2) and anaerobic threshold. Psychological measures included measures of mood, psychiatric symptoms, and neuropsychological functioning. This study demonstrated that 4 months of aerobic exercise training produced an overall 11.6% improvement in peak VO2 and a 13% increase in anaerobic threshold. In contrast, the Yoga and Waiting List control groups experienced no change in cardiorespiratory fitness. Other favorable physiological changes observed among aerobic exercise participants included lower cholesterol levels, diastolic blood pressure levels, and for subjects at risk for bone fracture, a trend toward an increase in bone mineral content. Although few significant psychological changes could be attributed to aerobic exercise training, participants in the two active treatment groups perceived themselves as improving on a number of psychological and behavioral dimensions. PMID:2768768

  3. Polysaccharides from Inonotus obliquus sclerotia and cultured mycelia stimulate cytokine production of human peripheral blood mononuclear cells in vitro and their chemical characterization.

    PubMed

    Xu, Xiangqun; Li, Juan; Hu, Yan

    2014-08-01

    Inonotus obliquus is an edible and medicinal mushroom to treat many diseases. In the present study, polysaccharides and fractions were isolated and purified by DEAE-52 and Sephadex G-200 chromatography from I. obliquus wild sclerotia, culture broth and cultured mycelia under submerged fermentation. The extracts and fractions could significantly induce the secretion of TNF-α, IFN-γ, IL-1β, and IL-2 in human peripheral blood mononuclear cells (PBMCs) and showed no toxicity to PBMCs. The stimulation effect of the six extracts and eight fractions on the four-cytokine production was dose-dependent. Sclerotial polysaccharides were more effective in the four-cytokine production at 150 μg/ml while exopolysaccharides and endopolysacchrides showed a much better effect on IL-1β production at 30 μg/ml. Purified fractions from exopolysaccharides and endopolysaccharides were more effective than the fraction from sclerotia in most cytokine production. These heteropolysaccharide-protein conjugates mainly contained glucose, galactose, and mannose. Protein content, molecular weight, monosaccharide molar ratio, and anomeric carbon configuration differed from each other and had effects on the cytokine induction activity of the polysaccharides to some extent. PMID:24867795

  4. Multicenter Comparison of ESP Culture System II with BACTEC 460TB and with Lowenstein-Jensen Medium for Recovery of Mycobacteria from Different Clinical Specimens, Including Blood

    PubMed Central

    Tortoli, Enrico; Cichero, Paola; Chirillo, M. Gabriella; Gismondo, M. Rita; Bono, Letizia; Gesu, Giampietro; Simonetti, M. Tullia; Volpe, Gisella; Nardi, Giampietro; Marone, Piero

    1998-01-01

    The recently developed ESP Culture System II (AccuMed, Chicago, Ill.) was compared with radiometric BACTEC 460TB (Becton Dickinson, Towson, Md.) and with Lowenstein-Jensen medium for recovery of mycobacteria from over 2,500 clinical specimens both of respiratory and nonrespiratory origin, including blood. The majority of the 219 mycobacterial isolates (129) belonged to the Mycobacterium tuberculosis complex, followed by 37 isolates of the Mycobacterium avium complex (MAC) and 53 isolates of eight other mycobacterial species. Rates of recovery obtained with BACTEC, ESP, and Lowenstein-Jensen medium were 89, 79, and 64%, respectively, with such differences being statistically significant. Different media and systems appeared to behave differently when the more frequently detected organisms were considered: M. tuberculosis complex isolates grew better with BACTEC, and MAC isolates grew better with ESP. An analysis of the combinations of Lowenstein-Jensen medium with BACTEC and with ESP did not reveal significant differences in recovery rates. With regard to the times needed for the detection of positive cultures, they were significantly longer on Lowenstein-Jensen medium (average, 28 days) than with the remaining two systems, between which there was no difference (average, 18 days). We conclude, therefore, that the ESP system, when used in combination with a solid medium, performs as well as the thoroughly validated radiometric BACTEC system and offers the advantages of full automation and absence of radioisotopes. PMID:9574709

  5. Aerobic Fitness for the Moderately Retarded.

    ERIC Educational Resources Information Center

    Bauer, Dan

    1981-01-01

    Intended for physical education teachers, the booklet offers ideas for incorporating aerobic conditioning into programs for moderately mentally retarded students. An explanation of aerobic fitness and its benefits is followed by information on initiating a fitness program with evaluation of height, weight, body fat, resting heart rate, and…

  6. Aerobic rice mechanization: techniques for crop establishment

    NASA Astrophysics Data System (ADS)

    Khusairy, K. M.; Ayob, H.; Chan, C. S.; Fauzi, M. I. Mohamed; Mohamad Fakhrul, Z. O.; Shahril Shah, G. S. M.; Azlan, O.; Rasad, M. A.; Hashim, A. M.; Arshad, Z.; E, E. Ibrahim; Saifulizan, M. N.

    2015-12-01

    Rice being the staple food crops, hundreds of land races in it makes the diversity of rice crops. Aerobic rice production was introduced which requires much less water input to safeguard and sustain the rice production and conserve water due to decreasing water resources, climatic changes and competition from urban and industrial users. Mechanization system plays an important role for the success of aerobic rice cultivation. All farming activities for aerobic rice production are run on aerobic soil conditions. Row seeder mechanization system is developed to replace conventional seeding technique on the aerobic rice field. It is targeted for small and the large scale aerobic rice farmers. The aero - seeder machine is used for the small scale aerobic rice field, while the accord - seeder is used for the large scale aerobic rice field. The use of this mechanization machine can eliminate the tedious and inaccurate seeding operations reduce labour costs and increases work rate. The machine is easy to operate and it can increase crop establishment rate. It reduce missing hill, increasing planting and crop with high yield can be produce. This machine is designed for low costs maintenance and it is easy to dismantle and assemble during maintenance and it is safe to be used.

  7. Skeletal Muscle Hypertrophy after Aerobic Exercise Training

    PubMed Central

    Konopka, Adam R.; Harber, Matthew P.

    2014-01-01

    Current dogma suggests aerobic exercise training has minimal effect on skeletal muscle size. We and others have demonstrated that aerobic exercise acutely and chronically alters protein metabolism and induces skeletal muscle hypertrophy. These findings promote an antithesis to the status quo by providing novel perspective on skeletal muscle mass regulation and insight into exercise-countermeasures for populations prone to muscle loss. PMID:24508740

  8. Aerobic Dancing--A Rhythmic Sport.

    ERIC Educational Resources Information Center

    Sorensen, Jacki

    Fitness programs now and in the future must offer built-in cardiovascular conditioning, variety, novelty, and change to meet the physical, mental, and emotional needs of our society. Aerobic dancing (dancing designed to train and strengthen the heart, lungs, and vascular system) is one of the first indoor group Aerobic exercise programs designed…

  9. Impact of positive chest X-ray findings and blood cultures on adverse outcomes following hospitalized pneumococcal lower respiratory tract infection: a population-based cohort study

    PubMed Central

    2013-01-01

    Background 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. Methods We studied a population-based multi-centre cohort of 705 adults hospitalized with LRTI and Streptococcus pneumoniae in LRT specimens or blood: 193 without pulmonary infiltrate or bacteraemia, 250 with X-ray confirmed pneumonia, and 262 with bacteraemia. We compared adverse outcomes in the three groups and used multiple regression analyses to adjust for differences in age, sex, comorbidity, and lifestyle factors. Results Patients with no infiltrate and no bacteraemia were of similar age but had more comorbidity than the other groups (Charlson index score ≥1: no infiltrate and no bacteraemia 81% vs. infiltrate without bacteraemia 72% vs. bacteraemia 61%), smoked more tobacco, and had more respiratory symptoms. In contrast, patients with a pulmonary infiltrate or bacteraemia had more inflammation (median C-reactive protein: no infiltrate and no bacteraemia 82 mg/L vs. infiltrate without bacteraemia 163 mg/L vs. bacteraemia 316 mg/L) and higher acute disease severity scores. All adverse outcomes increased from patients with no infiltrate and no bacteraemia to those with an infiltrate and to those with bacteraemia: Length of hospital stay (5 vs. 6 vs. 8 days); intensive care admission (7% vs. 20% vs. 23%); pulmonary complications (1% vs. 5% vs. 14%); and 30-day mortality (5% vs. 11% vs. 21%). Compared with patients with no infiltrate and no bacteraemia, the adjusted 30-day mortality rate ratio was 1.9 (95% confidence interval (CI) 0.9-4.1) in patients with an infiltrate without bacteraemia and 4.1 (95% CI 2.0-8.5) in bacteraemia patients. Adjustment for acute disease severity and inflammatory markers weakened these

  10. A novel co-culture model of the blood-retinal barrier based on primary retinal endothelial cells, pericytes and astrocytes.

    PubMed

    Wisniewska-Kruk, Joanna; Hoeben, Kees A; Vogels, Ilse M C; Gaillard, Pieter J; Van Noorden, Cornelis J F; Schlingemann, Reinier O; Klaassen, Ingeborg

    2012-03-01

    Loss of blood-retinal barrier (BRB) properties is an important feature in the pathology of diabetic macular edema (DME), but cellular mechanisms underlying BRB dysfunction are poorly understood. Therefore, we developed and characterized a novel in vitro BRB model, based on primary bovine retinal endothelial cells (BRECs). These cells were shown to maintain specific in vivo BRB properties by expressing high levels of the endothelial junction proteins occludin, claudin-5, VE-cadherin and ZO-1 at cell borders, and the specific pumps glucose transporter-1 (GLUT1) and efflux transporter P-glycoprotein (MDR1). To investigate the influence of pericytes and astrocytes on BRB maintenance in vitro, we compared five different co-culture BRB models, based on BRECs, bovine retinal pericytes (BRPCs) and rat glial cells. Co-cultures of BRECs with BRPCs and glial cells showed the highest trans-endothelial resistance (TEER) as well as decreased permeability of tracers after vascular endothelial growth factor (VEGF) stimulation, suggesting a major role for these cell types in maintaining barrier properties. To mimic the in vivo situation of DME, we stimulated BRECs with VEGF, which downregulated MDR1 and GLUT1 mRNA levels, transiently reduced expression levels of endothelial junctional proteins and altered their organization, increased the number of intercellular gaps in BRECs monolayers and influence the permeability of the model to differently-sized molecular tracers. Moreover, as has been shown in vivo, expression of plasmalemma vesicle-associated protein (PLVAP) was increased in endothelial cells in the presence of VEGF. This in vitro model is the first co-culture model of the BRB that mimicks in vivo VEGF-dependent changes occurring in DME. PMID:22200486

  11. Clinical Impact of Laboratory Implementation of Verigene BC-GN Microarray-Based Assay for Detection of Gram-Negative Bacteria in Positive Blood Cultures.

    PubMed

    Walker, Tamar; Dumadag, Sandrea; Lee, Christine Jiyoun; Lee, Seung Heon; Bender, Jeffrey M; Cupo Abbott, Jennifer; She, Rosemary C

    2016-07-01

    Gram-negative bacteremia is highly fatal, and hospitalizations due to sepsis have been increasing worldwide. Molecular tests that supplement Gram stain results from positive blood cultures provide specific organism information to potentially guide therapy, but more clinical data on their real-world impact are still needed. We retrospectively reviewed cases of Gram-negative bacteremia in hospitalized patients over a 6-month period before (n = 98) and over a 6-month period after (n = 97) the implementation of a microarray-based early identification and resistance marker detection system (Verigene BC-GN; Nanosphere) while antimicrobial stewardship practices remained constant. Patient demographics, time to organism identification, time to effective antimicrobial therapy, and other key clinical parameters were compared. The two groups did not differ statistically with regard to comorbid conditions, sources of bacteremia, or numbers of intensive care unit (ICU) admissions, active use of immunosuppressive therapy, neutropenia, or bacteremia due to multidrug-resistant organisms. The BC-GN panel yielded an identification in 87% of Gram-negative cultures and was accurate in 95/97 (98%) of the cases compared to results using conventional culture. Organism identifications were achieved more quickly post-microarray implementation (mean, 10.9 h versus 37.9 h; P < 0.001). Length of ICU stay, 30-day mortality, and mortality associated with multidrug-resistant organisms were significantly lower in the postintervention group (P < 0.05). More rapid implementation of effective therapy was statistically significant for postintervention cases of extended-spectrum beta-lactamase-producing organisms (P = 0.049) but not overall (P = 0.12). The Verigene BC-GN assay is a valuable addition for the early identification of Gram-negative organisms that cause bloodstream infections and can significantly impact patient care, particularly when resistance markers are detected. PMID:27098961

  12. Rapid Identification and Susceptibility Testing of Candida spp. from Positive Blood Cultures by Combination of Direct MALDI-TOF Mass Spectrometry and Direct Inoculation of Vitek 2

    PubMed Central

    Idelevich, Evgeny A.; Grunewald, Camilla M.; Wüllenweber, Jörg; Becker, Karsten

    2014-01-01

    Fungaemia is associated with high mortality rates and early appropriate antifungal therapy is essential for patient management. However, classical diagnostic workflow takes up to several days due to the slow growth of yeasts. Therefore, an approach for direct species identification and direct antifungal susceptibility testing (AFST) without prior time-consuming sub-culturing of yeasts from positive blood cultures (BCs) is urgently needed. Yeast cell pellets prepared using Sepsityper kit were used for direct identification by MALDI-TOF mass spectrometry (MS) and for direct inoculation of Vitek 2 AST-YS07 card for AFST. For comparison, MALDI-TOF MS and Vitek 2 testing were performed from yeast subculture. A total of twenty four positive BCs including twelve C. glabrata, nine C. albicans, two C. dubliniensis and one C. krusei isolate were processed. Applying modified thresholds for species identification (score ≥1.5 with two identical consecutive propositions), 62.5% of BCs were identified by direct MALDI-TOF MS. AFST results were generated for 72.7% of BCs directly tested by Vitek 2 and for 100% of standardized suspensions from 24 h cultures. Thus, AFST comparison was possible for 70 isolate-antifungal combinations. Essential agreement (minimum inhibitory concentration difference ≤1 double dilution step) was 88.6%. Very major errors (VMEs) (false-susceptibility), major errors (false-resistance) and minor errors (false categorization involving intermediate result) amounted to 33.3% (of resistant isolates), 1.9% (of susceptible isolates) and 1.4% providing 90.0% categorical agreement. All VMEs were due to fluconazole or voriconazole. This direct method saved on average 23.5 h for identification and 15.1 h for AFST, compared to routine procedures. However, performance for azole susceptibility testing was suboptimal and testing from subculture remains indispensable to validate the direct finding. PMID:25489741

  13. Nitrogen removal by Providencia rettgeri strain YL with heterotrophic nitrification and aerobic denitrification.

    PubMed

    Ye, Jun; Zhao, Bin; An, Qiang; Huang, Yuan-Sheng

    2016-09-01

    Providencia rettgeri strain YL shows the capability of nitrogen removal under sole aerobic conditions. By using isotope ratio mass spectrometry, (15)N-labelled N2O and N2 were detected in aerobic batch cultures containing [Formula: see text], [Formula: see text] or [Formula: see text]. Strain YL converted [Formula: see text], [Formula: see text] and [Formula: see text] to produce more N2O than N2 in the presence of [Formula: see text]. An (15)N isotope tracing experiment confirmed that the nitrogen removal pathway of strain YL was heterotrophic nitrification-aerobic denitrification. The optimal treatment conditions for nitrogen removal were pH of 8, C/N ratio of 12, temperature of 25°C and shaking speed of 105 rpm. A continuous aerobic bioreactor inoculated with strain YL was developed. With an influent [Formula: see text] concentration of 90-200 mg/L, the [Formula: see text] removal efficiency ranged from 80% to 97% and the total nitrogen removal efficiency ranged from 72% to 95%. The nitrogen balance in the continuous bioreactor revealed that approximately 35-52% of influent [Formula: see text] was denitrified aerobically to form gaseous nitrogen. These findings show that the P. rettgeri strain YL has potential application in wastewater treatment for nitrogen removal under sole aerobic conditions. PMID:26824874

  14. Nitrogen removal capability through simultaneous heterotrophic nitrification and aerobic denitrification by Bacillus sp. LY.

    PubMed

    Zhao, Bin; He, Yi Liang; Zhang, Xiao Fan

    2010-04-01

    The heterotrophic nitrification and aerobic denitrification capabilities of Bacillus sp. LY were investigated under the aerobic condition. The results indicate that Bacillus sp. LY is not only a heterotrophic nitrifier, but also an aerobic denitrifier. Experiments were carried out in an attempt to determine and quantify the contribution of heterotrophic nitrification and aerobic denitrification to total N removal. By taking the nitrogen balance under the culture condition of 41.1 mg/L of initial NH(4+)-N at a C/N ratio of 15 in 96 h, 8.0% of the initial NH(4)+-N still remained in the medium in the forms of hydroxylamine, nitrite, nitrate and organic N; 40.5% of NH(4+)-N was converted to biomass and 45.9% of NH(4+)-N was estimated to be finally removed in the formation of N2. This conversion of ammonium to N2 with the intermediate formation of N2O under the aerobic condition was confirmed by gas chromatography. Single step nitrogen removal by simultaneous heterotrophic nitrification and aerobic denitrification has great potential in wastewater treatment. PMID:20450115

  15. [Cardiovascular consequences of aerobic maneuvers].

    PubMed

    Trivelloni, Pierandrea; Berrettini, Umberto

    2010-10-01

    Gravitational (G) stress during aerobatics flights, both military and civilian, can suddenly incapacitate pilots in agile and supermaneuverable aircrafts. High +Gz stress, up to +9Gz, has two different physiological consequences: the first is the drop in head-level blood pressure that is proportional to the G load; the other, slightly delayed, is the blood pooling in the lower part of the body and the abdomen. This blood shift results in a decreased return of venous blood to the heart, decreased cardiac output, and decreased blood pressure, leading to a likely loss of consciousness. The natural countermeasure against the effects of high G stress is the baroreceptor reflex. The human physiological tolerance to the gravito-inertial forces developed in flight operations can be increased by physiological and technological means. PMID:21416842

  16. Selective production of interferon-alpha subtypes by cultured peripheral blood mononuclear cells and lymphoblastoid cell lines.

    PubMed Central

    Greenway, A L; Overall, M L; Sattayasai, N; Rowley, M J; Hertzog, P J; McMullen, G L; Cheetham, B F; Marzuki, S

    1992-01-01

    The biological significance of the existence of multiple interferon-alpha (IFN-alpha) subtypes is unknown but may represent a finely tuned mechanism whereby different subtypes are produced in response to different stimuli. To investigate the expression of individual IFN-alpha subtypes, polyclonal antipeptide antisera designed to react with all IFN-alpha subtypes, or with a particular subtype, IFN-alpha 2 or IFN-alpha 4, have been produced. In this study we demonstrate the utility of these antisera for the detection, using indirect immunofluorescence staining, of intracellular IFN-alpha produced by human peripheral blood mononuclear cells (PBMC) and lymphoblastoid cells. Secreted IFN-alpha was also investigated by bioassay and a sandwich radioimmunoassay (RIA), using two monoclonal antibodies (mAb) and specific for IFN-alpha 4. The PBMC were shown to produce IFN reactive with all three polyclonal antisera, after stimulation with Sendai virus. The lymphoblastoid cells also produced IFN, including IFN-alpha 2, but IFN-alpha 4 was not detected either intracellularly, by immunofluorescence, or in the medium, by sandwich RIA. The immunofluorescence studies also demonstrate that in the absence of viral stimulation IFN-alpha is found in the cytoplasm of PBMC and lymphoblastoid cells but not secreted in detectable levels. The finding that two lymphoblastoid cell lines do not produce the subtype IFN-alpha 4 raises important questions as to whether other cell lines and cell types produce IFN-alpha subtypes selectively, and whether individual IFN-alpha subtypes have different roles in human physiology and pathology. Images Figure 1 Figure 2 Figure 3 PMID:1537595

  17. Genome Sequence of "Pedosphaera parvula" Ellin514, an Aerobic Verrucomicrobial Isolate from Pasture Soil

    SciTech Connect

    Kant, Ravi; Van Passel, Mark W.J.; Palva, Airi; Lucas, Susan; Copeland, A; Lapidus, Alla L.; Glavina Del Rio, Tijana; Dalin, Eileen; Tice, Hope; Bruce, David; Goodwin, Lynne A.; Pitluck, Sam; Chertkov, Olga; Larimer, Frank W; Land, Miriam L; Hauser, Loren John; Brettin, Thomas S; Detter, J. Chris; Han, Cliff; De Vos, Willem M.; Janssen, Peter H.; Smidt, Hauke

    2011-01-01

    Pedosphaera parvula Ellin514 is an aerobically grown verrucomicrobial isolate from pasture soil. In contrast to the high abundance of members of Verrucomicrobia subdivision 3 based on molecular surveys in terrestrial environments, Ellin514 is one of the few cultured representatives of this group.

  18. Aerobic Biodegradation Kinetics And Mineralization Of Six Petrodiesel/Soybean-Biodiesel Blends

    EPA Science Inventory

    The aerobic biodegradation kinetics and mineralization of six petrodiesel/soybean-biodiesel blends (B0, B20, B40, B60, B80, and B100), where B100 is 100% biodiesel, were investigated by acclimated cultures. The fatty acid methyl esters (FAMEs) of biodiesel were found to undergo ...

  19. Bioaugmentation and enhanced formation of microbial granules used in aerobic wastewater treatment.

    PubMed

    Ivanov, Volodymyr; Wang, Xiao-Hui; Tay, Stephen Tiong-Lee; Tay, Joo-Hwa

    2006-04-01

    Microbial aggregates of an aerobic granular sludge can be used for the treatment of industrial or municipal wastewater, but their formation from a microbial activated sludge requires several weeks. Therefore, the aim of this research was the selection of microbial cultures to shorten the granule-forming period from several weeks to a few days. An enrichment culture with the ability to accelerate granulation was obtained by repeating the selection and batch cultivation of fast-settling microbial aggregates isolated from the aerobic granular sludge. Bacterial cultures of Klebsiella pneumoniae strain B and Pseudomonas veronii strain F, with self-aggregation indexes of 65 and 51%, respectively, and a coaggregation index of 58%, were isolated from the enrichment culture. A mixture of these strains with the activated sludge was used as an inoculum in an experimental sequencing batch reactor to start up an aerobic granulation process. Aerobic granules with a mean diameter of 446+/-76 microm were formed in an experiment after 8 days of cultivation, but microbial granules were absent in controls. Considering biosafety issues, K. pneumoniae strain B was excluded from further studies, but P. veronii strain F was selected for larger-scale testing. PMID:16091930

  20. Mass culture of photobacteria to obtain luciferase

    NASA Technical Reports Server (NTRS)

    Chappelle, E. W.; Picciolo, G. L.; Rich, E., Jr.

    1969-01-01

    Inoculating preheated trays containing nutrient agar with photobacteria provides a means for mass culture of aerobic microorganisms in order to obtain large quantities of luciferase. To determine optimum harvest time, growth can be monitored by automated light-detection instrumentation.

  1. Analysis on the Effect of Individualized Aerobic Exercise Intervention for Teenagers with Type 2 Diabetes

    PubMed Central

    Chun-qi, Zhao

    2015-01-01

    Objective: To investigate the intervention effect of individualized aerobic exercise on type 2 diabetes in teenagers. Method: To select 60 cases of teenager with type 2 diabetes in Zhoukou Hospital of Traditional Medicine in February 2013 to February 2014 as the research object, test all enrolled patients’ maximal oxygen and blood glucose fluctuation, and then give individualized aerobic exercise intervention, after 6 months intervention, compare the changes of patients’ indexes and evaluate the effect of individualized aerobic exercise intervention. Result: After the intervention, the patients’ plasma triglyceride and cholesterol content are significantly lower than before (P < 0.05); there’s no significant difference between high and low-density lipoprotein (P > 0.05). Moreover, the patients’ insulin and C-peptide level are significantly higher than those before intervention (P < 0.05); before intervention, their blood glucose and glycated hemogiobin level are higher than normal, after intervention, they are weakened, but there’s no significant difference (P > 0.05). The maximal oxygen uptake and different intensity of metabolic equivalents are higher than before, but there’s no significant difference (P > 0.05). Conclusion: For teenagers with type 2 diabetes, the implementation of individualized aerobic exercise intervention can effectively improve the patients’ lipid metabolism and cardio-pulmonary function, and effectively promote the insulin and C-peptide secretion, to provide scientific basis for effective control of blood glucose. PMID:26981161

  2. Fit women are not able to use the whole aerobic capacity during aerobic dance.

    PubMed

    Edvardsen, Elisabeth; Ingjer, Frank; Bø, Kari

    2011-12-01

    Edvardsen, E, Ingjer, F, and Bø, K. Fit women are not able to use the whole aerobic capacity during aerobic dance. J Strength Cond Res 25(12): 3479-3485, 2011-This study compared the aerobic capacity during maximal aerobic dance and treadmill running in fit women. Thirteen well-trained female aerobic dance instructors aged 30 ± 8.17 years (mean ± SD) exercised to exhaustion by running on a treadmill for measurement of maximal oxygen uptake (VO(2)max) and peak heart rate (HRpeak). Additionally, all subjects performed aerobic dancing until exhaustion after a choreographed videotaped routine trying to reach the same HRpeak as during maximal running. The p value for statistical significance between running and aerobic dance was set to ≤0.05. The results (mean ± SD) showed a lower VO(2)max in aerobic dance (52.2 ± 4.02 ml·kg·min) compared with treadmill running (55.9 ± 5.03 ml·kg·min) (p = 0.0003). Further, the mean ± SD HRpeak was 182 ± 9.15 b·min in aerobic dance and 192 ± 9.62 b·min in treadmill running, giving no difference in oxygen pulse between the 2 exercise forms (p = 0.32). There was no difference in peak ventilation (aerobic dance: 108 ± 10.81 L·min vs. running: 113 ± 11.49 L·min). In conclusion, aerobic dance does not seem to be able to use the whole aerobic capacity as in running. For well endurance-trained women, this may result in a lower total workload at maximal intensities. Aerobic dance may therefore not be as suitable as running during maximal intensities in well-trained females. PMID:22080322

  3. Effects of essential oils, yeast culture and malate on rumen fermentation, blood metabolites, growth performance and nutrient digestibility of Baluchi lambs fed high-concentrate diets.

    PubMed

    Malekkhahi, M; Tahmasbi, A M; Naserian, A A; Danesh Mesgaran, M; Kleen, J L; Parand, A A

    2015-04-01

    The experiment was conducted to evaluate the effects of dietary supplementation with a mixture of essential oils (MEO), yeast culture (YC) and malate on performance, nutrient digestion, rumen fermentation and blood metabolites of lambs fed high-concentrate growing diets. For this purpose, twenty Baluchi lambs (17.3 ± 0.5 kg body weight and 3 months old) were randomly assigned to four dietary treatments in a completely randomized design with five lambs per treatment. The treatment groups were as follows: (i) control: basal diet without any additive, (ii) basal diet plus 400 mg/day MEO (thymol, carvacrol, eugenol, limonene and cinnamaldehyde), (iii) basal diet with 4 g/day YC and (iv) basal diet plus 4 g/day malate. No differences between the dietary treatments were observed in dry matter intake, average daily gain or feed conversion ratio (p > 0.05). Compared with control and malate treatment, lambs fed MEO and YC had an improved crude protein digestibility (p < 0.05). Yeast culture significantly increased (p > 0.05) cell wall digestibility compared to the other treatments. No differences were observed between treatments with respect to nitrogen balance or ruminal pH and ammonia concentrations (p > 0.05). No differences were observed between treatments with respect to ruminal total volatile fatty acid concentration and molar proportions of acetate, butyrate and valerate. Molar proportion of propionate was higher (p < 0.05) for YC and malate compared to control and MEO. Plasma glucose concentration was higher (p < 0.05) in lambs fed YC and malate than in lambs fed the control or the MEO diet. Blood concentration of triglycerides significantly decreased when feeding the MEO and YC diets (p < 0.05). It was concluded that YC may be more useful as a feed additive for manipulation of rumen fermentation in lambs fed with high-concentrate diets than MEO and malate, because YC enhanced crude protein and cell wall digestibility, ruminal molar proportion

  4. Susceptibility Profile of Staphylococcus epidermidis and Staphylococcus haemolyticus Isolated from Blood Cultures to Vancomycin and Novel Antimicrobial Drugs over a Period of 12 Years.

    PubMed

    Pinheiro, Luiza; Brito, Carla Ivo; Pereira, Valéria Cataneli; Oliveira, Adilson; Bartolomeu, Ariane Rocha; Camargo, Carlos Henrique; Cunha, Maria Lourdes Ribeiro Souza

    2016-06-01

    The aim of this study was to evaluate the antimicrobial susceptibility profile of 85 Staphylococcus epidermidis and 84 Staphylococcus haemolyticus strains isolated from blood cultures to oxacillin, vancomycin, tigecycline, linezolid, daptomycin, and quinupristin/dalfopristin over a period of 12 years. S. epidermidis and S. haemolyticus isolated from blood cultures of inpatients, attended at a teaching hospital, were analyzed for the presence of the mecA gene and by SCCmec typing. The minimum inhibitory concentration (MIC) values of tigecycline, linezolid, daptomycin, quinupristin/dalfopristin, and vancomycin were determined. Isolates exhibiting vancomycin MICs of ≥2 μg/ml were typed by pulsed-field gel electrophoresis (PFGE). The rate of mecA positivity was 92.9% and 100% in S. epidermidis and S. haemolyticus, respectively. The most frequent SCCmec types were type III (53.2%) in S. epidermidis and type I (32.1%) in S. haemolyticus. All isolates were susceptible to linezolid and daptomycin, but 7.1% of S. haemolyticus and 2.3% of S. epidermidis isolates were resistant to tigecycline, and 1.2% each of S. haemolyticus and S. epidermidis were resistant and intermediately resistant to quinupristin/dalfopristin, respectively. S. epidermidis exhibited higher vancomycin MICs (40% with MIC of ≥2 μg/ml). Clonal typing of strains with vancomycin MIC of ≥2 μg/ml revealed the presence of different PFGE types of S. epidermidis and S. haemolyticus over a period of up to 4 years (2002-2004, 2005-2008, 2006-2009, 2010-2011). Despite the observation of a high prevalence of mecA, the clinical strains were fully susceptible to vancomycin and to the new drugs linezolid, daptomycin, tigecycline, and quinupristin/dalfopristin. The PFGE types with vancomycin MIC of ≥2 μg/ml exhibited a great diversity of SCCmec cassettes, demonstrating that S. epidermidis and S. haemolyticus may easily acquire these resistance-conferring genetic elements. PMID:26623676

  5. Effects of added chelated trace minerals, organic selenium, yeast culture, direct-fed microbials, and Yucca schidigera extract in horses. Part I: Blood nutrient concentration and digestibility.

    PubMed

    Gordon, M E; Edwards, M S; Sweeney, C R; Jerina, M L

    2013-08-01

    The objective of this study was to test the hypothesis that feed additives such as chelated minerals, organic Se, yeast culture, direct-fed microbials, and Yucca schidigera extract would improve nutrient digestibility when included in an equine diet. Horses (Quarter Horse geldings 4.5 to 16 yr of age; mean BW 522 kg ± 46 kg) were acclimated to 100% pelleted diets formulated with (ADD) and without (CTRL) commercially available sources of the aforementioned additives followed by a 14-d collection period of feces and urine. Chelated sources of Cu, Zn, Mn and Co were utilized versus sulfated forms, at a 100% replacement rate. No significant differences among apparent the digestibility of DM, ADF, or NDF (P= 0.665, P = 0.866, P = 0.747, respectively) were detected between dietary treatments. Likewise, no differences in apparent digestibility of Cu (P = 0.724), Zn (P = 0.256), Mn (P = 0.888), Co (P = 0.71), or Se (P = 0.588) were observed. No differences were observed in serum Cu, Mn, or Co concentrations between ADD and CTRL at acclimation or collection time points (P > 0.05). While no difference in serum Zn concentrations were observed between ADD and CTRL groups at acclimation (P > 0.05), they were statistically higher at the collection time period for horses consuming CTRL (P < 0.0001). Whole blood Se concentration was greater in the CTRL group versus the ADD group both at acclimation (P = 0.041) and collection (P = 0.005) time periods. In reference to time, serum Cu concentrations increased (P = 0.012) for animals consuming CTRL, but not ADD (P > 0.05). Serum Zn concentrations of horses consuming both ADD (P = 0.021) and CTRL (P < 0.0001) increased over time from acclimation to collection time points. No time differences (P > 0.05) were observed in serum Mn concentrations. Serum Co concentrations increased over time in horses consuming both ADD (P = 0.001) and CTRL (P = 0.021). From acclimation to collection, whole blood Se concentration increased for horses

  6. Blood Disorders

    MedlinePlus

    ... and protein. Over half of your blood is plasma. The solid part of your blood contains red blood cells, white blood cells and platelets. Blood disorders affect one or more parts of the blood and prevent ...

  7. Extensive Functional Evaluations to Monitor Aerobic Training in Becker Muscular Dystrophy: A Case Report

    PubMed Central

    Tramonti, Caterina; Rossi, Bruno; Chisari, Carmelo

    2016-01-01

    Low-intensity aerobic training seems to have positive effects on muscle strength, endurance and fatigue in Becker Muscular Dystrophy (BMD) patients. We describe the case of a 33-year old BMD man, who performed a four-week aerobic training. Extensive functional evaluations were executed to monitor the efficacy of the rehabilitative treatment. Results evidenced an increased force exertion and an improvement in muscle contraction during sustained exercise. An improvement of walk velocity, together with agility, endurance capacity and oxygen consumption during exercise was observed. Moreover, an enhanced metabolic efficiency was evidenced, as shown by reduced lactate blood levels after training. Interestingly, CK showed higher levels after the training protocol, revealing possible muscle damage. In conclusion, aerobic training may represent an effective method improving exercise performance, functional status and metabolic efficiency. Anyway, a careful functional assessment should be taken into account as a useful approach in the management of the disease’s rehabilitative treatment. PMID:27478558

  8. Velvet pad surface sampling of anaerobic and aerobic bacteria: an in vitro laboratory model.

    PubMed Central

    Raahave, D; Friis-Møller, A

    1982-01-01

    Velvet pads have been evaluated in an experimental, laboratory model, simulating intraoperative sampling of Staphylococcus epidermis, Escherichia coli and Bacteroides fragilis. After sampling, the pad was placed in a transport medium and kept in an anaerobic atmosphere, before being shaken and rinsed, followed by anaerobic and aerobic culture. This technique permitted quantitatively high recoveries of the test bacteria. Velvet pad sampling could be a measure to determine the density of aerobic and anaerobic bacteria during operation in an effort to predict the risk of postoperative wound sepsis. Images PMID:6757273

  9. Infection by HIV-1 blocked by binding of dextrin 2-sulphate to the cell surface of activated human peripheral blood mononuclear cells and cultured T-cells.

    PubMed Central

    Shaunak, S; Gooderham, N J; Edwards, R J; Payvandi, N; Javan, C M; Baggett, N; MacDermot, J; Weber, J N; Davies, D S

    1994-01-01

    1. Structural analogues of a sulphated polysaccharide, dextrin sulphate, were synthesized and tested for their ability to block infection by HIV-1. Using the T-cell lines, C8166 and HPB-ALL, and the laboratory adapted strains of HIV-1.MN, HIV-1.IIIb and HIV-1.RF, dextrin 2-sulphate (D2S) combined the best combination of high anti-HIV-1 activity (95% inhibitory concentration (IC95) = 230 nM) and low anticoagulant activity. It also blocked infection of activated peripheral blood mononuclear (PBMN) cells by five primary viral isolates at an IC95 of 230-3700 nM depending upon the primary viral isolate tested. 2. In saturation binding studies, [3H]-D2S bound to a cell surface protein on HPB-ALL cells in a specific and saturable manner with a Kd of 82 +/- 14 nM and a Bmax of 4.8 +/- 0.3 pmol/10(6) cells. It bound to other human T-cell lines in a similar manner. 3. There was very little binding of [3H]-D2S to freshly isolated PBMN cells (Bmax 0.18 +/- 0.03 pmol/10(6) cells) and these cells could not be infected by HIV-1. Culture of PBMN cells in lymphocyte growth medium (LGM) containing IL-2 did not significantly change the Bmax of [3H]-D2S. In contrast, PBMN cells which had been cultured with phytohaemagglutinin (PHA; 5 micrograms ml-1) for 72 h had a Bmax of [3H]-D2S binding of 7.2 +/- 0.1 pmol/10(6) cells and these cells could be infected by HIV-1. Removal of the PHA and further culture of the PBMN cells in LGM containing IL-2 resulted in a fall in the Bmax to 2.0 +/- 0.1 pmol/10(6) cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7812605

  10. Differences in Preseason Aerobic Fitness Screening in Professional and Pre-professional Modern Dancers.

    PubMed

    Bronner, Shaw; Codman, Emma; Hash-Campbell, Dana; Ojofeitimi, Sheyi

    2016-03-01

    The aerobic demands of today's dance repertoire warrant understanding of the current cardiorespiratory fitness of dancers. The purpose of this study was to compare aerobic fitness levels of professional and pre-professional modern dancers and determine change over time. A retrospective analysis of four groups, two professional, and two pre-professional, was conducted in preseason annual screens, occurring before the professional dancers' rehearsal period and the students' academic training. Resting (HRrest), peak (HRpeak), and recovery (HRrecov) heart rate, and blood pressure (BP) were compared in 577 dancers, using an accelerated 3-minute step test. Smoking, asthma, and aerobic and cross training rates between groups were also compared. A 4 (group) X 2 (gender) MANOVA design determined differences between groups and genders in all dependent variables (p < 0.05). Using a repeated measures ANOVA design, we compared a subgroup over 3 years and one pre-professional group over 4 years. There were differences between groups in systolic BP and all HR variables (p < 0.001). Professional dancers reflected better cardiorespiratory fitness than pre-professional dancers. There were differences between groups in aerobic and cross training activities but no differences in smoking incidence or asthma rates. Pre-professional dancers demonstrated improvement in aerobic fitness over time (p = 0.006) while professionals did not change. Professional dancers display better aerobic fitness, which may reflect their performance demands. Wellness programs appear to enhance fitness in pre-professional dance students over time. Additional aerobic training is recommended for pre-professional modern dance students to prepare them for the performance demands of a professional career. PMID:27025448

  11. Blood sugar test - blood

    MedlinePlus

    ... drink a certain amount of glucose ( oral glucose tolerance test ) How the Test will Feel When the ... a fasting blood glucose, HbA1c test , or glucose tolerance test , depending on your random blood glucose test ...

  12. The Energetics of Aerobic versus Anaerobic Respiration.

    ERIC Educational Resources Information Center

    Champion, Timothy D.; Schwenz, Richard W.

    1990-01-01

    Background information, laboratory procedures, and a discussion of the results of an experiment designed to investigate the difference in energy gained from the aerobic and anaerobic oxidation of glucose are presented. Sample experimental and calculated data are included. (CW)

  13. Neuromodulation of Aerobic Exercise—A Review

    PubMed Central

    Heijnen, Saskia; Hommel, Bernhard; Kibele, Armin; Colzato, Lorenza S.

    2016-01-01

    Running, and aerobic exercise in general, is a physical activity that increasingly many people engage in but that also has become popular as a topic for scientific research. Here we review the available studies investigating whether and to which degree aerobic exercise modulates hormones, amino acids, and neurotransmitters levels. In general, it seems that factors such as genes, gender, training status, and hormonal status need to be taken into account to gain a better understanding of the neuromodular underpinnings of aerobic exercise. More research using longitudinal studies and considering individual differences is necessary to determine actual benefits. We suggest that, in order to succeed, aerobic exercise programs should include optimal periodization, prevent overtraining and be tailored to interindividual differences, including neuro-developmental and genetically-based factors. PMID:26779053

  14. Aerobic Dance for Children: Resources and Recommendations.

    ERIC Educational Resources Information Center

    Wood, Denise A.

    1986-01-01

    Aerobic dance classes may be safe for older children, but are inappropriate for children in the fourth grade and under. Programs for these children should emphasize creativity. Resources for program development are given. (MT)

  15. Conditioning and Aerobics for Older Americans.

    ERIC Educational Resources Information Center

    Hansen, Joyce

    1980-01-01

    A class designed for the maintenance and gradual improvement of senior citizens' physical fitness includes relaxation training, flexibility and stretching exercises, interval training activities (designed as a link between less strenuous exercise and more strenuous activities), and aerobic exercises. (CJ)

  16. Cultured peripheral blood mast cells from chronic idiopathic urticaria patients spontaneously degranulate upon IgE sensitization: Relationship to expression of Syk and SHIP-2.

    PubMed

    Saini, Sarbjit S; Paterniti, Miya; Vasagar, Kavitha; Gibbons, Scott P; Sterba, Patricia M; Vonakis, Becky M

    2009-09-01

    Recently, signaling changes in the FcvarepsilonRI pathway involving inositol lipid phosphatases have been identified in the basophils of chronic idiopathic urticaria (CIU) subjects. Based on the profile of basophil FcvarepsilonRI-mediated histamine degranulation, we have segregated CIU subjects into two groups, CIU Responder (CIU R) or CIU Nonresponder (CIU NR). In the present study, we compared expression of SHIP-1, SHIP-2, and Syk protein to histamine release (HR) from mast cells (MC) cultured from the peripheral blood of CIU R, CIU NR, and normal subjects. The MC of CIU R donors contained significantly increased Syk and decreased SHIP-2 as compared to CIU NR (Syk: p=0.038, SHIP-2: p=0.038) and normals (Syk: p=0.042, SHIP-2: p=0.027). Spontaneous HR from CIU donors was increased two-fold compared to normals (p=0.04). In summary, our results suggest a possible predilection for urticarial MC to spontaneously degranulate upon IgE sensitization contributing to the increased pruritus associated with CIU. PMID:19477690