scholarly journals Effects of Decontamination Methods and Culture Conditions on Viability of Mycobacterium ulcerans in the BACTEC System

1998 ◽  
Vol 36 (2) ◽  
pp. 402-408 ◽  
Author(s):  
J. C. Palomino ◽  
F. Portaels
Keyword(s):  
Geographic Origin ◽  
Egg Yolk ◽  
Polymyxin B ◽  
Culture Conditions ◽  
Mycobacterium Ulcerans ◽  
Clinical Specimens ◽  
Geographic Origins ◽  
Primary Cultivation ◽  
Bactec System ◽  
Detrimental Impact

We used the BACTEC system to evaluate the effects of several decontamination methods and the addition of antibiotics on the viability of Mycobacterium ulcerans. The effects of polyoxyethylene stearate or egg yolk as supplements were also evaluated to determine their impact on the growth of M. ulcerans. Strains of different geographic origins were subjected to Petroff, reversed Petroff, oxalic acid, and mild HCl treatments. After treatment, the viability of each strain was assessed in the BACTEC system. All of the decontamination methods tested adversely affected bacterial viability. Treatment with mild HCl gave the best results, allowing better growth rates with some strains and causing a delay in growth with others, depending on the geographic origin of the strain. A mixture of polymyxin B, amphotericin B, nalidixic acid, trimethoprim, and azlocillin did not significantly inhibit growth. Supplementing BACTEC medium with egg yolk markedly improved the recovery of M. ulcerans following the use of each of the decontamination methods. Our findings demonstrate a detrimental impact on the viability of M. ulcerans by all of the decontamination methods currently in common use. This explains, at least in part, the difficulty often experienced in cultivating this organism from clinical specimens. Egg yolk should be added to enhance the rate of successful primary cultivation of M. ulcerans in the BACTEC system.

scholarly journals Primary cultivation: factors affecting contamination and Mycobacterium ulcerans growth after long turnover time of clinical specimens

2014 ◽  
Vol 14 (1) ◽  
Author(s):  
Martin W Bratschi ◽  
Miriam Bolz ◽  
Leticia Grize ◽  
Sarah Kerber ◽  
Jacques C Minyem ◽  
...  
Keyword(s):  
Turnover Time ◽  
Mycobacterium Ulcerans ◽  
Factors Affecting ◽  
Clinical Specimens ◽  
Primary Cultivation

scholarly journals Genotyping Mycobacterium ulcerans and Mycobacterium marinum by Using Mycobacterial Interspersed Repetitive Units

2005 ◽  
Vol 187 (5) ◽  
pp. 1639-1647 ◽  
Author(s):  
Pieter Stragier ◽  
Anthony Ablordey ◽  
Wayne M. Meyers ◽  
Françoise Portaels
Keyword(s):  
Tandem Repeats ◽  
Central Africa ◽  
Mycobacterium Ulcerans ◽  
Pathogenic Species ◽  
Missing Link ◽  
Potential Value ◽  
Related Organism ◽  
Geographic Origins ◽  
West And Central Africa ◽  
Unique Genotype

ABSTRACT A novel category of variable tandem repeats (VNTR) called mycobacterial interspersed repetitive units (MIRUs) has been identified for Mycobacterium ulcerans (n = 39), M. marinum (n = 27), and one related organism. Fifteen MIRU loci were identified in the genome of M. marinum and were used to genotype M. ulcerans, M. marinum, and an M. marinum-like organism that is considered a possible missing link between M. marinum and M. ulcerans. Seven MIRU loci were polymorphic, and locus-specific PCRs for four of these loci differentiated seven M. ulcerans genotypes, four M. marinum genotypes, and a unique genotype for the missing link organism. The seven M. ulcerans genotypes were related to six different geographic origins of isolates. All isolates from West and Central Africa, including old and recent isolates, belonged to the same genotype, emphasizing the great spatiotemporal homogeneity among African isolates. Unlike the M. ulcerans genotypes, the four M. marinum genotypes could not be clearly related to the geographic origins of the isolates. According to MIRU-VNTR typing, all M. ulcerans and M. marinum isolates of American origin were closely related, suggesting a common American ancestor for these two pathogenic species on the American continents. MIRU typing has significant potential value for discriminating between reoccurrence and reinfection for M. ulcerans disease.

Effect of Oxygen on Growth of Mycobacterium ulcerans in the BACTEC System

1998 ◽  
Vol 36 (11) ◽  
pp. 3420-3422 ◽  
Author(s):  
J. C. Palomino ◽  
A. M. Obiang ◽  
L. Realini ◽  
W. M. Meyers ◽  
F. Portaels
Keyword(s):  
Primary Culture ◽  
Oxygen Concentration ◽  
Oxygen Tension ◽  
Mycobacterium Ulcerans ◽  
Clinical Samples ◽  
Low Oxygen ◽  
Bactec System ◽  
Effect Of Oxygen ◽  
Low Oxygen Concentration

The effect of low oxygen concentration on the growth of 15 strains of Mycobacterium ulcerans was evaluated in the BACTEC system. Reduced oxygen tension enhanced the growth of M. ulcerans, suggesting that this organism has a preference for microaerobic environments. Application of this observation may improve rates of isolation of M. ulcerans in primary culture from clinical samples and promote isolation of the bacterium from environmental sources.

EXTENDED STUDY OF SERRATIA IN A DIAGNOSTIC BACTERIOLOGY LABORATORY

1966 ◽  
Vol 12 (1) ◽  
pp. 99-103 ◽  
Author(s):  
R. M. Hotz ◽  
V. R. Dowell Jr.
Keyword(s):  
Polymyxin B ◽  
Biochemical Tests ◽  
Diagnostic Laboratory ◽  
Extended Study ◽  
Clinical Specimens ◽  
Hospital Acquired Infections ◽  
Noticeable Increase ◽  
Routine Identification ◽  
Hospital Acquired ◽  
Antibiotic Agents

The incidence of Serratia in clinical material received in a general hospital diagnostic laboratory was studied. Initially the morphologic features, biochemical characteristics, chromogenesis, and antibiotic susceptibility of 21 isolates were examined. On the basis of these studies certain cultural and biochemical tests were selected for routine identification of Serratia.Serratia isolated from various types of clinical specimens received during a 4-year period (1960–1964) were recorded. Most of the Serratia were nonchromogenic except on a special medium. The cultures were resistant to a number of antibiotic agents, particularly penicillin, polymyxin B, and the tetracyclines. A higher incidence of Serratia in sputum than in throat and nasopharyngeal cultures was observed. There was a noticeable increase in the overall incidence of Serratia isolates over the 4-year period. This may be a reflection of hospital acquired infections.

scholarly journals Distribution of carbapenemases and efflux pump in carbopenems-resistance Acinetobacter baumannii

2016 ◽  
Author(s):  
Zhu-Qiang Qiu ◽  
Li-Jing Zhu ◽  
Pan-Fei Hou
Keyword(s):  
Acinetobacter Baumannii ◽  
Efflux Pump ◽  
Polymyxin B ◽  
Clinical Specimens ◽  
Expression Difference ◽  
Serious Infections ◽  
The World ◽  
Agar Dilution ◽  
First Time ◽  
Antibiotic Genes

Acinetobacter baumannii has emerged as an important pathogen related to serious infections and nosocomial outbreaks around the world. The aim of this study was to detect the distribution of carbapenemases and efflux pump in carbopenems-resistance Acinetobacter baumannii(CRAB). In this study, 100 isolates of CRAB were collected from clinical specimens. Agar dilution was conducted to determine the minimum inhibitory concentrations (MICs) to 15 kinds of antibiotic. Genes of carbapenemases and efflux pumps were amplified by PCR. The expression difference of pump genes was also analyzed by real-time PCR between CRAB and carbopenems- sensitive Acinetobacter baumannii (CSAB). We found that most antibiotics, including aminoglycosides, fluoroquinolones and cephalosporins showed high MIC values in CRAB. While, all isolates were sensitive to polymyxin B. Among CRAB, 54, 32 and 16 isolates were positive for SHV-12，PER-1 and TEM-1, respectively. 86 isolates were positive for OXA-23. 55, 33 and 5 isolates carried adeB, adeJ and adeE genes. The expression level of adeB in CRAB was ten times higher than that in CSAB. Moreover, isolates with single adeE gene were detected for the first time in Acinetobacter baumannii.

scholarly journals 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

1998 ◽  
Vol 36 (5) ◽  
pp. 1378-1381 ◽  
Author(s):  
Enrico Tortoli ◽  
Paola Cichero ◽  
M. Gabriella Chirillo ◽  
M. Rita Gismondo ◽  
Letizia Bono ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Mycobacterium Avium ◽  
Culture System ◽  
Solid Medium ◽  
Mycobacterial Species ◽  
Becton Dickinson ◽  
Clinical Specimens ◽  
Lowenstein Jensen ◽  
Bactec System ◽  
The Times

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 theMycobacterium 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.

scholarly journals Isolation of Blood-Borne Mycobacterium avium by Using the Nonradioactive BACTEC 9000 MB System and Comparison with a Solid-Culture System

1998 ◽  
Vol 36 (12) ◽  
pp. 3703-3706 ◽  
Author(s):  
Véronique Jacomo ◽  
Didier Musso ◽  
Marie-José Gevaudan ◽  
Michel Drancourt
Keyword(s):  
Mycobacterium Avium ◽  
Polymyxin B ◽  
Automated System ◽  
Contamination Rate ◽  
Detection Time ◽  
Solid Culture ◽  
Conventional Procedure ◽  
Solid Culture Medium ◽  
Bactec System ◽  
Blood Specimens

We conducted a 12-month prospective study comparing two approaches to the detection of Mycobacterium avium in the blood of human immunodeficiency virus type 1-infected patients, namely, a lytic centrifugation system combined with Middlebrook solid culture medium (the conventional procedure) and the nonradiometric BACTEC 9000 MB system. Species identification relied on 16S rRNA probe hybridization and cell wall fatty acids chromatography. M. avium was isolated in 17 of 345 (5%) blood specimens by the BACTEC 9000 MB automated system and in 14 of 345 (4%) blood specimens by the conventional procedure (nonsignificant, χ2 test). Detection time was 16 ± 6 days by the BACTEC 9000 MB automated system and 27 ± 3 days by the conventional procedure (P < 0.001, Student t test). Non-M. avium mycobacteria were not recovered during the study period. Contamination rate was 8% (30 specimens) by the BACTEC 9000 MB system and 0% by the conventional procedure, indicating the necessity of using an antibiotic mixture (PANTA, consisting of polymyxin B, amphotericin B, nalidixic acid, trimethoprim, and azlocillin). Working time was 1 min 30 s by the BACTEC 9000 MB system and 8 min by the conventional procedure, which was 1.8 times more expensive than the BACTEC system. Use of the BACTEC 9000 MB system increased the sensitivity of M. avium detection and reduced detection time in blood culture.

Induction of plasminogen activator inhibitor (PAI) by lipopolysaccharide (LPS)

1987 ◽  
Author(s):  
A M DOSNE ◽  
F DUBOR ◽  
L CHEDID
Keyword(s):  
Endothelial Cells ◽  
Plasminogen Activator ◽  
Plasminogen Activator Inhibitor ◽  
Polymyxin B ◽  
Culture Conditions ◽  
Clot Lysis ◽  
Human Endothelial Cells ◽  
Lysis Time ◽  
Binding Factor ◽  
Activator Inhibitor

It has been shown that, under culture conditions, human endothelial cells synthetize plasminogen activator inhibitor (PAI) which neutralize urokinase and tissue plasminogen activator.Treatment of human endothelial cells with LPS (50 ngto 10 μg/ml) from S. enteritidis resulted in a dose-dependent increase in PAI production.Fibrinoenzymographic analysis showed that incubation of supernatantfrom LPS-treated cells with urokinase of low and high mol. w. (33.000 and 55.000) led to disappearance of the two urokinase lytic bands and formation of high mol. w. complexes (Mr 93.000 and 107.000). The mol. w. of the urokinase binding factor was calculated to be near 50.000. Polymyxin B and colimycin could suppress this effect of LPS. Injection of LPS (30 ng-30 yg/kg in the rat led to a considerable decrease in the fibrinolytic activity of plasma euglobulins which clot lysis time was prolonged from 55 up to morethan 240 min. This hypofibrinolytic state was associated with PAI detected in euglobulins and in plasma.Large complexes (Mr 80.000-105.000) were formed between exogenous urokinase of low and high mol. w. mixed with post LPS plasma or euglobulins. Polymyxin B and Colimycin could prevent the hypofibrinolytic response to low doses of LPS. These results suggest thatPAI generation in endotoxemia could be due in part to the direct effect of LPS on endothelium.

scholarly journals In Vitro Susceptibility of Mycobacterium ulcerans to Clarithromycin

1998 ◽  
Vol 42 (8) ◽  
pp. 2070-2073 ◽  
Author(s):  
F. Portaels ◽  
H. Traore ◽  
K. De Ridder ◽  
W. M. Meyers
Keyword(s):  
Bacterial Infections ◽  
Geographic Origin ◽  
Surgical Excision ◽  
Etiologic Agent ◽  
Mycobacterium Ulcerans ◽  
World Health ◽  
In Vitro Susceptibility ◽  
Strong Activity

ABSTRACT Buruli ulcer (BU), caused by Mycobacterium ulcerans, was recently recognized by the World Health Organization as an important emerging disease. While antimycobacterial therapy is often effective for the earliest nodular or ulcerative lesions, medical management of BU lesions in patients presenting for treatment is usually disappointing, leaving wide surgical excision the only alternative. Advanced ulcerated lesions of BU rarely respond to antimycobacterial agents; however, perioperative administration of such drugs may prevent relapses or disseminated infections. Clarithromycin possesses strong activity in vitro and in vivo against most nontuberculous mycobacteria. In this study we determined the antimycobacterial activity of this drug in vitro against 46 strains ofM. ulcerans isolated from 11 countries. The MIC of clarithromycin was determined at pH 6.6 (on 7H11 agar) and at pH 7.4 (on Mueller-Hinton agar). The MICs ranged from 0.125 to 2 μg/ml at pH 6.6 and from <0.125 to 0.5 μg/ml at pH 7.4. For the majority of the strains, geographic origin did not play a significant role. Thirty-eight strains (83%) were inhibited by 0.5 μg/ml at pH 7.4. These MICs are below peak therapeutic concentrations of clarithromycin obtainable in blood. These results suggest that clarithromycin is a promising drug both for the treatment of early lesions of M. ulcerans and for the prevention of hematogenous dissemination of the etiologic agent during and after surgery. Studies should be initiated to evaluate the effects of clarithromycin in combination with ethambutol and rifampin on M. ulcerans both in vitro and in experimentally infected mice. Multidrug regimens containing clarithromycin may also help control the secondary bacterial infections sometimes seen in BU patients, most importantly osteomyelitis.

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