scholarly journals Different culture media containing methyldopa for melanin production by Cryptococcus species

2011 ◽  
Vol 44 (5) ◽  
pp. 591-594 ◽  
Author(s):  
Ralciane de Paula Menezes ◽  
Mário Paulo Amante Penatti ◽  
Reginaldo dos Santos Pedroso
Keyword(s):  
Sunflower Seed ◽  
Minimal Medium ◽  
Culture Media ◽  
Melanin Production ◽  
Pharmaceutical Tablet ◽  
Mueller Hinton ◽  
Cryptococcus Species ◽  
Mueller Hinton Agar ◽  
The Media ◽  
Niger Seed

INTRODUCTION: Melanin production by species of Cryptococcus is widely used to characterize C. neoformans complex in mycology laboratories. This study aims to test the efficacy of methyldopa from pharmaceutical tablet as a substrate for melanin production, to compare the production of melanin using different agar base added with methyldopa, and to compare the melanin produced in those media with that produced in Niger seed agar and sunflower seed agar by C. neoformans, C. laurentii, and C. albidus. Two isolates of each species, C. neoformans, C. laurentii, and C. albidus, and one of Candida albicans were used to experimentally detect conditions for melanin production. METHODS: The following media were tested: Mueller-Hinton agar (MHA), brain and heart infusion agar (BHIA), blood agar base (BAB), and minimal medium agar (MMA), all added with methyldopa, and the media Niger seed agar (NSA) and sunflower seed agar (SSA). RESULTS: All isolates grew in most of the culture media after 24h. Strains planted on media BAB and BHIA showed growth only after 48h. All isolates produced melanin in MMA, MHA, SSA, and NSA media. CONCLUSIONS: Methyldopa in the form pharmaceutical tablet can be used as a substrate for melanin production by Cryptococcus species; minimal medium plus methyldopa was more efficient than the BAB, MHA, and BHIA in the melanin production; and NSA and SSA, followed by MMA added with methyldopa, were more efficient than other media studied for melanin production by all strains studied.

scholarly journals Method for evaluating broth culture media: application to Haemophilus

1978 ◽  
Vol 8 (5) ◽  
pp. 520-524
Author(s):  
A W Brinkley ◽  
T W Huber
Keyword(s):  
Nicotinamide Adenine Dinucleotide ◽  
Research Laboratory ◽  
Culture Media ◽  
Nicotinamide Adenine ◽  
Broth Culture ◽  
Mueller Hinton ◽  
The Media ◽  
Growth Promoting ◽  
Mueller Hinton Broth

A method was devised to test the growth-promoting ability of a broth medium. The "dilute to extinction" method determines the inoculum required to develop heavy turbidity in a broth with overnight incubation. A statistical method using Poisson distribution was used to show that a single Haemophilus cell can develop heavy turbidity in an optimal broth. The dilute to extinction method was used to evaluate the shelf life of stored media, to titrate the growth factor requirements of Haemophilus, and to evaluate the use of purified hemin and nicotinamide adenine dinucleotide in a broth medium for the growth of Haemophilus. Of the media tested, the most suitable formulation was Mueller-Hinton broth supplemented with 10 microgram of hemin and 10 microgram of nicotinamide adenine dinucleotide per ml. The dilute to extinction method appears to be especially useful in the development of broth media for fastidious organisms. The method could also be used to assure the quality of other broth media which are required to support the growth of small inocula in the clinical or research laboratory.

scholarly journals Dextranomer/Hyaluronic Acid Copolymer (Dx/HA) Has no Effect on Bacterial Growth in Culture Media With or Without Antibiotic Discs

2020 ◽  
Vol 9 (2) ◽  
Author(s):  
Karabulut R ◽  
Doğruman-Al F ◽  
Türkyılmaz Z ◽  
Sönmez K ◽  
Demirel F ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Bacterial Growth ◽  
Culture Media ◽  
Acid Copolymer ◽  
Mueller Hinton ◽  
Mueller Hinton Agar ◽  
Agar Media ◽  
Inhibition Zones ◽  
Made In

Introduction: Endoscopic treatment of vesicoureteral refl ux (VUR) by subureteral injection of biocompatible polymers is an established treatment option for refl ux. Dextranomer/hyaluronic acid copolymer(Dx/HA) has gained wide popularity for treating VUR. We decided to investigate the antibacterial activity of Dx/HA and its interaction with antibiotics in in-vitro conditions. Materials and Methods: Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and Proteus mirabilis suspensions were inoculated into Mueller-Hinton agar media and 30 μl of Dx/HA was inoculated in 5 mm diameter pits and the plates were incubated at 37°C for 24 hours. At the end of the incubation period, inhibition zones around the discs were measured. Expansion of the inhibition zones towards the pits which contained Dx/HA was considered as synergism. Dx/ HA was inoculated into pits made in Mueller-Hinton agar medium without antibiotic discs but containing suspensions of bacteria. These media were incubated under the same circumstances and same measurements were done. All experimental procedures were performed twice. Increase in bacterial zone diameters for ≥ 5 mm was inoculated was regarded as signifi cant for each agent. Results: Dx/HA caused no difference in bacterial growth either with or without antibiotic discs as determined by inhibition zones in the culture media. Conclusion : Dx/ha will not contribute to UTI if it is used for the treatment of VUR in cases either with or without infection.

scholarly journals Applicability and performance of EUCAST’s rapid antimicrobial susceptibility testing (RAST) on primarily sterile body fluids in blood culture bottles in laboratory routine with total lab automation

2021 ◽  
Author(s):  
Jasmin Kaur Jasuja ◽  
Stefan Zimmermann ◽  
Irene Burckhardt
Keyword(s):  
Blood Culture ◽  
Susceptibility Testing ◽  
Reference Method ◽  
Body Fluids ◽  
E Coli ◽  
Mueller Hinton ◽  
Mueller Hinton Agar ◽  
And Performance ◽  
Sterile Body Fluids ◽  
Better Than

AbstractOptimisation of microbiological diagnostics in primarily sterile body fluids is required. Our objective was to apply EUCAST’s RAST on primarily sterile body fluids in blood culture bottles with total lab automation (TLA) and to compare results to our reference method Vitek2 in order to report susceptibility results earlier. Positive blood culture bottles (BACTEC™ Aerobic/Anaerobic/PEDS) inoculated with primarily sterile body fluids were semi-automatically subcultured onto Columbia 5% SB agar, chocolate agar, MacConkey agar, Schaedler/KV agar and Mueller-Hinton agar. On latter, cefoxitin, ampicillin, vancomycin, piperacillin/tazobactam, meropenem and ciprofloxacin were added. After 6 h, subcultures and RAST were imaged and MALDI-TOF MS was performed. Zone sizes were digitally measured and interpreted following RAST breakpoints for blood cultures. MIC values were determined using Vitek2 panels. During a 1-year period, 197 Staphylococcus aureus, 91 Enterococcus spp., 38 Escherichia coli, 11 Klebsiella pneumoniae and 8 Pseudomonas aeruginosa were found. Categorical agreement between RAST and MIC was 96.5%. Comparison showed no very major errors, 2/7 (28.6%) and 1/7 (14.3%) of major errors for P. aeruginosa and meropenem and ciprofloxacin, 1/9 (11.1%) for K. pneumoniae and ciprofloxacin, 4/69 (7.0%) and 3/43 (5.8%) for Enterococcus spp. and vancomycin and ampicillin, respectively. Minor errors for P. aeruginosa and meropenem (1/8; 12.8%) and for E. coli and ciprofloxacin (2/29; 6.5%) were found. 30/550 RAST measurements were within area of technical uncertainty. RAST is applicable and performs well for primarily sterile body fluids in blood culture bottles, partially better than blood-based RAST. Official EUCAST evaluation is needed.

scholarly journals Recording the Presence of Peanibacillus larvae larvae Colonies on MYPGP Substrates Using a Multi-Sensor Array Based on Solid-State Gas Sensors

Sensors ◽  
2021 ◽  
Vol 21 (14) ◽  
pp. 4917
Author(s):  
Beata Bąk ◽  
Jakub Wilk ◽  
Piotr Artiemjew ◽  
Jerzy Wilde
Keyword(s):  
Gas Sensors ◽  
Culture Media ◽  
Laboratory Diagnosis ◽  
Baseline Correction ◽  
American Foulbrood ◽  
Sensor Signal ◽  
Sodium Pyruvate ◽  
Mueller Hinton ◽  
Regeneration Phase ◽  
Mueller Hinton Broth

American foulbrood is a dangerous disease of bee broods found worldwide, caused by the Paenibacillus larvae larvae L. bacterium. In an experiment, the possibility of detecting colonies of this bacterium on MYPGP substrates (which contains yeast extract, Mueller-Hinton broth, glucose, K2HPO4, sodium pyruvate, and agar) was tested using a prototype of a multi-sensor recorder of the MCA-8 sensor signal with a matrix of six semiconductors: TGS 823, TGS 826, TGS 832, TGS 2600, TGS 2602, and TGS 2603 from Figaro. Two twin prototypes of the MCA-8 measurement device, M1 and M2, were used in the study. Each prototype was attached to two laboratory test chambers: a wooden one and a polystyrene one. For the experiment, the strain used was P. l. larvae ATCC 9545, ERIC I. On MYPGP medium, often used for laboratory diagnosis of American foulbrood, this bacterium produces small, transparent, smooth, and shiny colonies. Gas samples from over culture media of one- and two-day-old foulbrood P. l. larvae (with no colonies visible to the naked eye) and from over culture media older than 2 days (with visible bacterial colonies) were examined. In addition, the air from empty chambers was tested. The measurement time was 20 min, including a 10-min testing exposure phase and a 10-min sensor regeneration phase. The results were analyzed in two variants: without baseline correction and with baseline correction. We tested 14 classifiers and found that a prototype of a multi-sensor recorder of the MCA-8 sensor signal was capable of detecting colonies of P. l. larvae on MYPGP substrate with a 97% efficiency and could distinguish between MYPGP substrates with 1–2 days of culture, and substrates with older cultures. The efficacy of copies of the prototypes M1 and M2 was shown to differ slightly. The weighted method with Canberra metrics (Canberra.811) and kNN with Canberra and Manhattan metrics (Canberra. 1nn and manhattan.1nn) proved to be the most effective classifiers.

scholarly journals 1455. Potential Mechanisms of Cefiderocol MIC Increase in Enterobacterales in In Vitro Resistance Acquisition Studies

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S730-S730
Author(s):  
Yoshinori Yamano ◽  
Rio Nakamura ◽  
Miki Takemura ◽  
Roger Echols
Keyword(s):  
Iron Transport ◽  
Resistance Mechanisms ◽  
Altered Expression ◽  
Carbapenem Resistant ◽  
Mueller Hinton ◽  
Two Component ◽  
Mueller Hinton Agar ◽  
Related Proteins

Abstract Background Cefiderocol (CFDC) is a novel siderophore, iron-chelating cephalosporin, which is transported into bacteria via iron transporters. CFDC has potent in vitro and in vivo activity against all aerobic Gram-negative bacteria, including carbapenem-resistant strains. To date, clinical isolates with cefiderocol MIC >4 µg/mL have been found infrequently, in which the presence of a few β-lactamases or altered iron transport was found. We investigated potential new mechanisms causing CFDC MIC increases in non-clinical studies. Methods The mutation positions were determined by whole genome sequencing using four K. pneumoniae mutants including two KPC producers and one NDM producer that had shown CFDC MIC increases in previous in vitro resistance-acquisition studies. The mutant strains were obtained at the frequency of 10-7 to < 10-8 by spreading bacteria on standard Mueller‒Hinton agar medium containing CFDC at concentrations of 10× MIC, with or without apo-transferrin (20 μg/mL). CFDC MIC was determined by broth microdilution using iron-depleted cation-adjusted Mueller-Hinton broth based on Clinical and Laboratory Standards Institute guidelines. The emergence of MIC increase mutants was also assessed by in vitro chemostat models under humanized plasma pharmacokinetic exposures of CFDC. Results The possible resistance mechanisms were investigated. Mutation of baeS or envZ, sensors of two-component regulation systems, were found in three or two mutants among the tested four isolates, respectively, and caused the MIC to increase by 4–32-fold. The altered expression level of specific genes by the baeS or envZ mutation could affect CFDC susceptibility, but the specific genes have not been identified. In addition, the mutation of exbD, an accessory protein related to iron transport, was identified in one case and caused the MIC to increase by >8-fold. In vitro chemostat studies using two isolates (one NDM producer and one KPC producer) showed no resistance acquisition during 24-hour exposure. Table. Overview of mutation emergence in five isolates of K. pneumoniae Conclusion The mutation of two-component regulation systems (BaeSR and OmpR/EnvZ) and iron transport-related proteins were shown to be possible mechanisms causing CFDC MIC increases, but these mutants did not appear under human exposures. Disclosures Yoshinori Yamano, PhD, Shionogi & Co., Ltd. (Employee) Rio Nakamura, BSc, Shionogi & Co., Ltd. (Employee) Miki Takemura, MSc, Shionogi & Co., Ltd. (Employee) Roger Echols, MD, Shionogi Inc. (Consultant)

scholarly journals 1457. Serial Passage of Enterobacteriaceae to Explore Development of Carbapenem Resistance

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S731-S731
Author(s):  
Yosef Nissim ◽  
Douglas Slain ◽  
P Rocco LaSala
Keyword(s):  
Susceptibility Testing ◽  
Klebsiella Oxytoca ◽  
Carbapenem Resistance ◽  
Serial Passage ◽  
Resistant Isolate ◽  
Mueller Hinton ◽  
The Us ◽  
Zones Of Inhibition ◽  
Mueller Hinton Agar ◽  
The Impact

Abstract Background Carbapenems are broad-spectrum antibacterials that have seen increased usage for the Enterobacteriales family in recent years. While carbapenem usage has been associated with increased antibacterial resistance, there is currently a lack of data comparing the risk of reduced susceptibility selection by the two most commonly used carbapenems in the US, ertapenem (ERT) and meropenem (MER). We conducted a novel serial passage experiment with clinical isolates of Enterobacteriales to assess the impact of repeated exposure to ERT or MER on phenotypic susceptibility patterns. Methods Non-duplicate clinical Enterobacteriales isolates were selected randomly for inclusion. Antimicrobial susceptibility testing was performed by CLSI disc diffusion methods. Standardized suspensions of isolates were plated on Mueller-Hinton agar, and ERT (10mcg) and MER (10mcg) discs applied. Zones of inhibition were measured and recorded after 16-18 hours incubation. Growth from the innermost zone of inhibition around each disc was used to prepare subsequent suspensions for serial susceptibility testing. This process would be repeated daily for 10 days. Each subsequent serially-passaged isolated was tested against both ERT and MER. Daily zones of inhibition were measured and interpreted. Baseline & final susceptibilities were determined by automated methods (Vitek 2). Results Seventeen Enterobacteriaceae isolates were selected, including: Klebsiella pneumoniae (n=11), Klebsiella oxytoca (n=2), Escherichia coli (n=1), Morganella morganii (n=1), and Enterobacter cloacae (n=2). Despite a greater degree of reductions in zones of inhibition with repeated ERT exposure (vs MER), the overall 10 day trends were not found to be significant different (P=0.529). Resistance developed to ERT in six isolates compared to one MER-resistant isolate (P = 0.053). E. cloacae was the only species to show a significant change between drugs (P=0.010). Two of three isolates that developed reduced zone changes > 10mm to MER were initially exposed to ERT on an earlier plate. Conclusion This novel experiment identified the development of some nonsignificant reductions in susceptibility with ERT after serial exposure. Results from this pilot study should encourage larger well-designed studies in this area. Disclosures All Authors: No reported disclosures

scholarly journals The Roles of Porous Coral Sands in Initial Enrichment of Ammonia-Oxidizing Bacteria

2007 ◽  
Vol 7 ◽  
pp. 525-532
Author(s):  
Qing Guo ◽  
Zao-he Wu ◽  
Ming-liang Qian ◽  
Binhe Gu
Keyword(s):  
Culture Media ◽  
Potassium Dihydrogen Phosphate ◽  
N:P Ratio ◽  
Ammonium Nitrogen ◽  
Dihydrogen Phosphate ◽  
Ammonia Oxidizing Bacteria ◽  
Potassium Dihydrogen ◽  
Conversion Rates ◽  
Calcium Source ◽  
The Media

The purpose of this study was to investigate the roles of coral sands in the enrichment and isolation of ammonium-oxidizing bacteria (AOB). We hypothesized that the porous coral sands provided additional surface area and nutrients for the growth of periphytic AOB. In the present study, an orthogonal test was designed to compare the AOB conversion rates of ammonium-nitrogen (NH4+N) to nitrite-nitrogen (NO2--N) among various combinations of culture media. Results showed that the conversion of NH4+N to NO2--N increased significantly when the coral sands were added, implying that coral sands were beneficial to the growth of AOB. Additions of potassium dihydrogen phosphate (KH2PO4) or sodium bicarbonate (NaHCO3) to the media became unnecessary when coral sands were used, but the addition of KH2PO4was needed when the molar nitrogen to phosphorus (N:P) ratio reached 10 in the enrichment media using calcium carbonate (CaCO3) powder as a calcium source.

Biochemical markers for pregnancy in the spent culture medium of in vitro produced bovine embryos

2021 ◽  
Author(s):  
Gabriela de Oliveira Fernandes ◽  
Marcella Pecora Milazzotto ◽  
Andrei Antonioni Guedes Fidelis ◽  
Taynan Stonoga Kawamoto ◽  
Ligiane de Oliveira Leme ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Biochemical Markers ◽  
Culture Medium ◽  
Culture Media ◽  
Ionization Mass ◽  
Metabolic Profiles ◽  
Bovine Embryos ◽  
The Media

Abstract The present study aimed to identify biomarkers to assess the quality of in vitro produced (IVP) bovine embryos in the culture media. IVP embryos on Day (D) 5 of development were transferred to individual drops, where they were maintained for the last 48 h of culture. Thereafter, the medium was collected and the embryos were transferred to the recipients. After pregnancy diagnosis, the media were grouped into the pregnant and nonpregnant groups. The metabolic profiles of the media were analyzed via electrospray ionization mass spectrometry, and the concentrations of pyruvate, lactate, and glutamate were assessed using fluorimetry. The spectrometric profile revealed that the media from embryos from the pregnant group presented a higher signal intensity compared to that of the nonpregnant group; the ions 156.13 Da [M + H]+, 444.33 Da [M + H]+, and 305.97 Da [M + H]+ were identified as biomarkers. Spent culture medium from expanded blastocysts (Bx) that established pregnancy had a greater concentration of pyruvate (p = 0.0174) and lesser concentration of lactate (p = 0.042) than spent culture medium from Bx that did not establish pregnancy. Moreover, pyruvate in the culture media of Bx can predict pregnancy with 90.9% sensitivity and 75% specificity. In conclusion, we identified markers in the culture media that helped in assessing the most viable IVP embryos with a greater potential to establish pregnancy.

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