Antifungal activity was tested against three Aspergillus species associated with stonebrood disease (A. flavus ATCC9170, A. fumigatus ATCC10894, and A. niger ATCC10549) from Fraunhofer strain collection (STO20519-STO29521) using CLSI M51-A parameters [72 (link)]. The Aspergillus strains were cultivated on PDA (VWR International, Radnor, PA, USA) for 7 days at 25 °C. The spores were washed by 5–6 mL of 0.05% Tween-80 (Sigma-Aldrich, St. Louis, MO, USA) and filtered through three layers of miracloth to remove the hyphal structures. The inoculum was adjusted to OD530 = 0.14–0.46 based on spore morphology [73 (link)]. The inoculum was spread evenly over the surface of Mueller–Hinton agar (Carl Roth, Karlsruhe, Germany) using a sterile cotton swab. The depth of the medium was consistent in each Petri dish (4 mm). The extracts from the liquid T. purpureogenus strains cultures (Figure 1 ) were diluted with methanol to 10 mg/mL, and 25 µL of the crude extract was applied to 6 mm cellulose disks and left in the laminar flow cabinet for 30 min to dry. Methanol was used as a control. The disks were placed on the inoculated media and pressed down with sterile forceps. The zones of inhibition were evaluated in triplicate after incubation at 35 °C for 24 h.
Mueller hinton agar (mha)
Manufactured by Carl Roth
Sourced in Germany
Mueller–Hinton agar is a standard culture medium used for antimicrobial susceptibility testing. It provides the necessary nutrients and growth conditions for the cultivation of a wide range of bacteria.
Automatically generated - may contain errors
6 protocols using mueller hinton agar (mha)
1
Antifungal Activity Evaluation of Talaromyces Extracts
2
Viable Cell Enumeration in Biofilms
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To determine the number of viable cells by plate count (CFU), the non-adherent bacteria were removed by aspiration and each sample was cut into two equally sized pieces. One piece was used for histology and the other was transferred to 1 mL PBS (pH 7.4) and vortexed for 1 min to homogenize the biofilm. Next, the bacterial suspension was diluted and 100 µL of each dilution was plated on Mueller–Hinton agar (Carl Roth, Karlsruhe, Germany), followed by incubation overnight at 37 °C. Colonies were then counted and final counts calculated, taking the dilution factor into consideration.
3
Antimicrobial Susceptibility Testing Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antimicrobial susceptibility tests were performed using the Vitek 2 system (bioMérieux, Nürtingen, Germany) and the test card AST-P608, according to manufacturer’s instructions and guidelines of the European Committee on Antimicrobial Susceptibility Testing (EUCAST).
In addition, MICs for Trimethoprim/Sulfamethoxazole and Trimethoprim were determined using MIC Test Strips (bestbion dx, Köln, Germany/Liofilchem, Roseto degli Abruzzi, Italy), i.e., an agar diffusion technique that uses stripes impregnated with a gradient of an antibiotic compound in order to allow direct reading of the MIC. MIC Test Strips were used according to manufacturer’s instructions on Mueller Hinton agar (Roth, Karlsruhe, Germany).
In addition, MICs for Trimethoprim/Sulfamethoxazole and Trimethoprim were determined using MIC Test Strips (bestbion dx, Köln, Germany/Liofilchem, Roseto degli Abruzzi, Italy), i.e., an agar diffusion technique that uses stripes impregnated with a gradient of an antibiotic compound in order to allow direct reading of the MIC. MIC Test Strips were used according to manufacturer’s instructions on Mueller Hinton agar (Roth, Karlsruhe, Germany).
4
Antibiotic Resistance Profiling of Vibrio parahaemolyticus
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The three V. parahaemolyticus strains and the reference strain Escherichia (E.) coli ATCC 25922 (strain LMG8223 from the Belgian Coordinated Collection of Microorganisms BCCM) were tested for antibiotic resistance using the disc diffusion method, in compliance with the Clinical and Laboratory Standards Institute (CLSI) M45 guidelines for Vibrio spp. A total of 13 different antibiotics (all purchased from Oxoid Limited, UK) were tested: ampicillin (10 µg), amoxicillin-clavulanate (20/10 µg), ampicillin-sulbactam (10/10), piperacillin (100 µg), cefotaxime (30 µg), ceftazidime (30 µg), amikacin (30 µg), gentamicin (10 µg), tetracycline (30 µg), ciprofloxacin (5 µg), levofloxacin (5 µg), ofloxacin (5 µg) and trimethoprim-sulfamethoxazole (1.25/23.75 µg). Briefly, the bacteria were grown on Mueller–Hinton Agar (MHA, Carl Roth, Germany) plates after which a direct colony suspension was prepared in a 0.85% NaCl solution, and turbidity was adjusted to 0.5 McFarland standard. The MHA plates were inoculated with 100 µL of this suspension which was spread with a sterile triangle rod. Plates were allowed to dry for 5 to 10 min prior to applying the antibiotic disks onto the agar with sterile tweezers. The plates were incubated inverted at 35°C for 16 ± 2 h. The reference strain E. coli ATCC 25922 was used as a control to monitor the accuracy of the disk diffusion tests.
5
Standardized Microbial Culturing Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Test microorganisms—Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, and Candida albicans P37037—strains were provided by the culture collection of the Microbiology Laboratory, Alexandru Ioan Cuza University of Iasi. The methicillin-resistant Staphylococcus aureus (MRSA) isolate was kindly provided by Med. biol. PhD Simona Matiut from Praxis Clinical Laboratory, Iasi, Romania, and included in the same microbial culture collection mentioned above, with the following accession number: prx-MRSA-2018.
All strains were stored in 15% glycerol stocks at −80 °C. Prior to experiments, S. aureus and E. coli strains were transferred to Mueller–Hinton agar (MHA, Liofilchem, Roseto degli Abruzzi, Italy) and C. albicans on Sabouraud dextrose agar (SDA, Carl Roth, Karlsruhe, Germany) and incubated at 37 °C. Subsequently, 10 mL of Mueller–Hinton broth (MHB, Scharlau, Barcelona, Spain) and 15 mL of Sabouraud dextrose broth (SDB, Carl Roth, Germany) were inoculated with one representative colony of each test organism taken from MHA or SDA, cultured overnight (37 °C, 190 rpm and 130 rpm, respectively) and used as source of inoculum for further experiments.
All strains were stored in 15% glycerol stocks at −80 °C. Prior to experiments, S. aureus and E. coli strains were transferred to Mueller–Hinton agar (MHA, Liofilchem, Roseto degli Abruzzi, Italy) and C. albicans on Sabouraud dextrose agar (SDA, Carl Roth, Karlsruhe, Germany) and incubated at 37 °C. Subsequently, 10 mL of Mueller–Hinton broth (MHB, Scharlau, Barcelona, Spain) and 15 mL of Sabouraud dextrose broth (SDB, Carl Roth, Germany) were inoculated with one representative colony of each test organism taken from MHA or SDA, cultured overnight (37 °C, 190 rpm and 130 rpm, respectively) and used as source of inoculum for further experiments.
6
Antimicrobial Susceptibility Testing of Vibrio
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sixteen antimicrobial agents (Table 4
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!