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

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.

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A Study Evaluating Conventional and Automated System of Culture for Isolation of Mycobacteria from Smear Negative Tuberculosis Samples with Special Reference to MPT 64 Antigen

Journal of Evidence Based Medicine and Healthcare ◽

10.18410/jebmh/2021/76 ◽

2021 ◽

Vol 8 (07) ◽

pp. 391-395

Author(s):

Rehena Sarkar ◽

Debalina Das

Keyword(s):

Predictive Value ◽

Gold Standard ◽

Standard Technique ◽

Antigen Detection ◽

Automated System ◽

Contamination Rate ◽

Detection Time ◽

Tuberculosis Complex ◽

Biochemical Tests ◽

Smear Negative

BACKGROUND India is among one of the major tuberculosis (TB) endemic countries of the world. The Centers for Disease Control and Prevention (CDC) guidelines mandate early species identification of M. tuberculosis complex as an effective countermeasure. The conventional culture method, which is the ‘gold-standard’ technique of mycobacterial isolation is time consuming, while the newer, automated, liquid medium-based culture systems like BACT / ALERT 3D have considerably shorter detection time and greater sensitivity. Other than isolation it is also important to differentially identify Mycobacterium tuberculosis complex (MTBC) from nontuberculous mycobacteria (NTM). As MPT 64 antigen is specific for MTBC and can be detected with considerable accuracy it can be used as a tool to differentiate between MTBC and NTM. METHODS 200 samples (sputum, cerebrospinal fluid-CSF, pleural fluid etc.) in total were collected from clinically suspected smear negative patients. Each sample was inoculated both in L-J media and BACT / ALERT 3D system. Those samples which showed growth were further differentiated into MTBC and NTM both by conventional biochemical tests and MPT64 antigen detection kit test. RESULTS Out of the 200 samples, 30 produced growths. Lowenstein-Jensen (L-J) media detected 12 (40 %) and BACT / ALERT 3D system detected 26 (86.67 %) of the isolates. Among the 30 isolates, 18 (60 %) were MTBC and 12 (40 %) were NTM. Furthermore, L-J media detected 44.4 % and BACT / ALERT 3D system detected 88.9 % out of 18 MTBC isolates while among the 12 NTM isolates L-J media detected 33.33 % and BACT / ALERT 3D system detected 83.33 %. Mean detection time for MTBC was 46.5 days by L-J media and 20.8 days by BACT / ALERT 3D system. Mean detection time for NTM was 24.5 days by L-J media and 9.86 days by BACT / ALERT 3D system. Contamination rate in this study was 12 % in L-J media and 3 % in BACT / ALERT 3D system. We also found that for sensitivity, specificity, positive predictive value, negative predictive value of MPT64 antigen detection kit test was 100 % in our study considering biochemical tests as gold standard. CONCLUSIONS BACT / ALERT 3D system has a very good isolation rate and shorter mean detection time compared to L-J media even from smear negative samples. Also, MPT-64 antigen detection kit test is a very viable option to differentiate between MTBC and NTM. KEYWORDS MTBC, NTM, BACT / Alert 3D, MPT 64 Antigen

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Use of BACTEC MGIT 960 for Recovery of Mycobacteria from Clinical Specimens: Multicenter Study

Journal of Clinical Microbiology ◽

10.1128/jcm.37.11.3578-3582.1999 ◽

1999 ◽

Vol 37 (11) ◽

pp. 3578-3582 ◽

Cited By ~ 94

Author(s):

Enrico Tortoli ◽

Paola Cichero ◽

Claudio Piersimoni ◽

M. Tullia Simonetti ◽

Giampietro Gesu ◽

...

Keyword(s):

Oxygen Sensor ◽

Solid Medium ◽

Automated System ◽

Contamination Rate ◽

Mycobacterial Species ◽

Solid Media ◽

Mycobacterium Xenopi ◽

Significant Difference ◽

Lowenstein Jensen ◽

Automatic Systems

The BACTEC MGIT 960 instrument is a fully automated system that exploits the fluorescence of an oxygen sensor to detect growth of mycobacteria in culture. Its performance was compared to those of the radiometric BACTEC 460 instrument and egg-based Lowenstein-Jensen medium. An identical volume of sample was inoculated in different media, and incubation was carried out for 6 weeks with the automatic systems and for 8 weeks on solid media. A total of 2,567 specimens obtained from 1,631 patients were cultured in parallel. Mycobacteria belonging to nine different taxa were isolated by at least one of the culture systems, with 75% of them being represented byMycobacterium tuberculosis complex. The best yield was obtained with the BACTEC 460 system, with 201 isolates, in comparison with 190 isolates with the BACTEC MGIT 960 system and 168 isolates with Lowenstein-Jensen medium. A similar but not significant difference was obtained when the most-represented organisms, the M. tuberculosis complex, Mycobacterium xenopi, and theMycobacterium avium complex, were analyzed separately and when combinations of a solid medium with the BACTEC MGIT 960 system and with the BACTEC 460 system were considered. The shortest times to detection were obtained with the BACTEC MGIT 960 system (13.3 days); 1.5 days earlier than that with the BACTEC 460 system (14.8 days) and 12 days earlier than that with Lowenstein-Jensen medium (25.6 days). The BACTEC MGIT 960 system had a contamination rate of 10.0%, intermediate between those of the radiometric system (3.7%) and the egg-based medium (17.0%). We conclude, therefore, that the BACTEC MGIT 960 system is a fully automated, nonradiometric instrument that is suitable for the detection of growth of tuberculous and other mycobacterial species and that is characterized by detection times that are even shorter than that of the “gold standard,” the BACTEC 460 system. The contamination rate was higher than that for the radiometric BACTEC 460 system and needs to be improved.

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Recovery of Mycobacterium avium-M. intracellulare from blood specimens by using the routine BACTEC 6B blood culture system.

Journal of Clinical Microbiology ◽

10.1128/jcm.27.2.348-349.1989 ◽

1989 ◽

Vol 27 (2) ◽

pp. 348-349 ◽

Cited By ~ 2

Author(s):

R Horn ◽

A Laszlo ◽

H G Robson

Keyword(s):

Blood Culture ◽

Mycobacterium Avium ◽

Culture System ◽

Blood Culture System ◽

Blood Specimens

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Temporary immersion biorreators: efficient technique for the propagation of the ‘Pircinque’ strawberry

Revista Brasileira de Fruticultura ◽

10.1590/0100-29452019102 ◽

2019 ◽

Vol 41 (1) ◽

Author(s):

Samila Silva Camargo ◽

Leo Rufato ◽

Maicon Magro ◽

André Luiz Kulkamp de Souza

Keyword(s):

Liquid Medium ◽

Culture Medium ◽

Culture Media ◽

Time Factors ◽

Efficient Technique ◽

Control Treatment ◽

Solid Culture ◽

Temporary Immersion ◽

Solid Culture Medium

Abstract The in vitro propagation technique via temporary immersion bioreactors is a tool that, through the culture in a liquid medium, allows an increase in the efficiency of seedling production. Several researches with the strawberry crop have shown greater efficiency of the system compared to the conventional process of micropropagation in solid medium. In this sense, the objective herein was to establish a protocol of multiplication and rooting of the ‘Pircinque’ strawberry, in temporary immersion bioreactors. Two distinct and independent studies were carried out, characterized by the multiplication and rooting stages of strawberry explants, newly introduced and registered in Brazil. Two culture media (MS and KNOP) were studied and, as a control treatment, the growth of the explants in solid culture medium was evaluated with the addition of 5 g L-1 of agar. Different immersion times of the culture medium were explored: five or eight times a day, for 15 minutes. The study was composed of the culture medium and immersion time factors, as well as the control (solid) treatment. It was verified that the use of temporary immersion bioreactors system is an efficient technique for the multiplication and rooting of explants of strawberry cv. Pircinque, when compared to the conventional method of micropropagation with the use of solid culture medium, making it possible to optimize the production of seedlings in biofactories. The MS liquid medium, in contact with explants of ‘Pircinque’ strawberry five times a day, increased the growth of the aerial part and the root system.

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Immunopotentiating Activity of the Water-soluble Lignin Rich Fraction Prepared from LEM—The Extract of the Solid Culture Medium ofLentinus edodesMycelia—

Bioscience Biotechnology and Biochemistry ◽

10.1271/bbb.61.1909 ◽

1997 ◽

Vol 61 (11) ◽

pp. 1909-1912 ◽

Cited By ~ 29

Author(s):

Yoshiki Yamamoto ◽

Hiroyuki Shirono ◽

Keiko Kono ◽

Yasuhiro Ohashi

Keyword(s):

Culture Medium ◽

Water Soluble ◽

Solid Culture ◽

Solid Culture Medium ◽

Soluble Lignin

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Intracellular Biosynthesis of Fluorescent CdSe Quantum Dots inBacillus subtilis:A Strategy to Construct Signaling Bacterial Probes for Visually Detecting Interaction BetweenBacillus subtilisandStaphylococcus aureus

Microscopy and Microanalysis ◽

10.1017/s1431927615015548 ◽

2015 ◽

Vol 22 (1) ◽

pp. 13-21 ◽

Cited By ~ 5

Author(s):

Zheng-Yu Yan ◽

Xiao-Xia Ai ◽

Yi-Long Su ◽

Xin-Ying Liu ◽

Xiao-Hui Shan ◽

...

Keyword(s):

Quantum Dots ◽

Culture Medium ◽

Imaging Mass Spectrometry ◽

Cdse Quantum Dots ◽

Mass Spectrometry Detection ◽

New Paradigm ◽

Solid Culture ◽

Bacterial Interaction ◽

Solid Culture Medium ◽

Good Uniformity

AbstractIn this work, fluorescentBacillus subtilis(B. subtilis) cells were developed as probes for imaging applications and to explore behaviorial interaction betweenB. subtilisandStaphylococcus aureus(S. aureus). A novel biological strategy of coupling intracellular biochemical reactions for controllable biosynthesis of CdSe quantum dots by livingB. subtiliscells was demonstrated, through which highly luminant and photostable fluorescentB. subtiliscells were achieved with good uniformity. With the help of the obtained fluorescentB. subtiliscells probes,S. aureuscells responded to co-culturedB. subtilisand to aggregate. The degree of aggregation was calculated and nonlinearly fitted to a polynomial model. Systematic investigations of their interactions implied thatB. subtiliscells inhibit the growth of neighboringS. aureuscells, and this inhibition was affected by both the growth stage and the amount of surroundingB. subtiliscells. Compared to traditional methods of studying bacterial interaction between two species, such as solid culture medium colony observation and imaging mass spectrometry detection, the procedures were more simple, vivid, and photostable due to the efficient fluorescence intralabeling with less influence on the cells’ surface, which might provide a new paradigm for future visualization of microbial behavior.

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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

Journal of Clinical Microbiology ◽

10.1128/jcm.36.5.1378-1381.1998 ◽

1998 ◽

Vol 36 (5) ◽

pp. 1378-1381 ◽

Cited By ~ 34

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.

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STUDIES ON CERTAIN GROWTH PARAMETERS OF DIFFERENT PROBIOTIC STRAINS

Journal of Engineering Studies and Research ◽

10.29081/jesr.v23i4.70 ◽

2017 ◽

Vol 23 (4) ◽

pp. 19-24

Author(s):

DUMITRA RĂDUCANU ◽

ANA-MARIA GEORGESCU

Keyword(s):

Nutrient Medium ◽

Growth And Development ◽

Culture Medium ◽

Growth Parameters ◽

Probiotic Bacteria ◽

Solid Culture ◽

Solid Culture Medium ◽

Probiotic Strains ◽

Direct Counting ◽

Better Than

The researchers found that probiotics contain microorganisms belonging to genus: Streptococcus, Lactobacillus, Bacillus, Aspergillus, Saccharomyces, Enterococcus, Pediococcus, enzymes (lactoperoxidase, gluconase, nonspecific enzymes) and rumen extracts. In this study, commercial probiotic bacteria known as "Linex" were used as samples. Cultural characteristics of these probiotic bacteria have been isolated and studied. It has been found that solid culture medium (nutritional gelose) favored the growth and development of bifidobacteria better than the liquid nutrient medium (nutrient broth). Thus, the number of bifidobacteria resulting from direct counting with Thoma chamber was of 7890 cells.

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