Moraxella catarrhalis

Manufactured by American Type Culture Collection

Sourced in Canada, United States

Moraxella catarrhalis is a species of Gram-negative, aerobic bacteria that can be cultured and studied using laboratory equipment. The species is commonly found in the upper respiratory tract of humans.

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7 protocols using moraxella catarrhalis

Antibiotic Susceptibility Profiling

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Broth microdilution minimum inhibitory concentration (MIC) testing was performed in 96-well microtiter plates according to Clinical and Laboratory Standards Institute (CLSI, formerly NCCLS) guideline M7-A7.¹⁴ MIC values were determined for E. coli (ATCC 25922), E. coli tolC mutant, Enterococcus faecalis (ATCC 29212), Haemophilus influenzae (ATCC 49766), Moraxella catarrhalis (ATCC 25238), P. aeruginosa (ATCC 47085), P. aeruginosa PAO200 (efflux pump mutant), P. aeruginosa hypersensitive strain (ATCC 35151), Staphylococcus aureus (ATCC 29213), and Streptococcus pneumonia (ATCC 49619).
Time-kill studies were performed using E. faecalis and S. pneumoniae for compound BT_03F04 and S. aureus and S. pneumoniae for compound BT_04B09 based on the MIC assay results, according to CLSI document M26-A.¹⁵ Growth media was Brain Heart Infusion and Trypticase Soy Broth from Remel (Lenexa, KS) with or without 3% lysed horse blood (Hemostat Laboratories, Dixon, CA).

Hu Y., Guerrero E., Keniry M., Manrrique J, & Bullard J.M. (2015). Identification of Chemical Compounds That Inhibit the Function of Glutamyl-tRNA Synthetase from Pseudomonas aeruginosa. Journal of biomolecular screening, 20(9), 1160-1170.

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Microbiological Susceptibility Testing

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Broth microdilution MIC testing was carried out in 96-well microtiter plates according to Clinical Laboratory Standards Institute guideline M7-A7 [26 ]. MIC values were determined for E. coli (ATCC 25922), E. coli tolC mutant, Enterococcus faecalis (ATCC 29212), Haemophilus influenzae (ATCC 49766), Moraxella catarrhalis (ATCC 25238), P. aeruginosa (ATCC 47085), P. aeruginosa PAO200 (efflux pump mutant), P. aeruginosa hypersensitive strain (ATCC 35151), S. aureus (ATCC 29213), and S. pneumonia (ATCC 49619) from the American Type Culture Collection (Manassas, VA). For quality control (QC) of MIC determination and culture purity, MICs were determined for antibiotics specific for each bacterial strain. The antibiotic and concentration range for each bacteria was: E. coli (ampicillin, 0.125-128 μg/ml), E. faecalis (vancomycin 0.125-128 μg/ml), H. influenzae (ampicillin, 0.03125-32 μg/ml), M. catarrhalis (ampicillin, 0.015625-16 μg/ml), P. aeruginosa (ampicillin, 0.125-128 μg/ml), S. aureus (oxacillin, 0.03125-32 μg/ml), S. pneumoniae (penicillin, 0.0625-64 μg/ml).

Hu Y., Palmer S.O., Munoz H, & Bullard J.M. (2015). High Throughput Screen Identifies Natural Product Inhibitor of Phenylalanyl-tRNA Synthetase from Pseudomonas aeruginosa and Streptococcus pneumoniae. Current drug discovery technologies, 11(4), 279-292.

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Bacterial Strain Acquisition and Characterization

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Oligonucleotides were purchased from the Integrated DNA Technologies (Coralville, IA). All other materials were purchased from Sigma Aldrich (St. Louis, MO) or Fisher Scientific (Pittsburg, PA). DNA sequencing was carried out at the Howard Hughes Medical Institute (HHMI) laboratory at The University of Texas – Pan American and Functional Bioscience (Madison, WI). Radioactive isotopes were from PerkinElmer (Waltham, MA). Moraxella catarrhalis (ATCC 25238), Enterococcus faecalis (ATCC 29212), Streptococcus pneumonia (ATCC 49619), Escherichia coli (ATCC 25922), Staphylococcus aureus (ATCC 29213), Haemophilus influenza (ATCC 49766), Pseudomonas aeruginosa (ATCC 47085) were from the American Type Culture Collection (Manassas, VA). E. coli tolC mutant, P. aeruginosa PAO200 (efflux pump mutant) and P. aeruginosa hypersensitive strain (ATCC 35151) were a kind gift from Urs Ochsner (Crestone Pharma-Boulder CO). Human mitochondrial PheRS (hmPheRS) was prepared as described [21 (link)]. Chemical compounds were from Prestwick Chemical (Illkirch, France).

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Diverse Bacterial Strain Collection for Research

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P. aeruginosa (ATCC 25619 and clinical isolates), Bordetella bronchiseptica (ATCC 4617, ATCC 10580), Escherichia coli (ATCC 25922, ATCC 700973), Burkholderia cenocepacia (ATCC 25608 and clinical isolates), Acinetobacter lwoffii (ATCC 17925), Acinetobacter baumannii (ATCC 19606), Moraxella catarrhalis (ATCC 8176), Bacillus subtilis (ATCC 6633), and S. aureus (ATCC 29213 and clinical isolates) were purchased from Cedarlane (Burlington, ON, Canada) or obtained from the Clinical Microbiology Laboratory of Memorial Hospital (Sudbury, ON, Canada). For experimentation, the strains were inoculated onto Mueller Hinton II agar plates and incubated for 24 h at 37°C.

Coccimiglio J., Alipour M., Jiang Z.H., Gottardo C, & Suntres Z. (2016). Antioxidant, Antibacterial, and Cytotoxic Activities of the Ethanolic Origanum vulgare Extract and Its Major Constituents. Oxidative Medicine and Cellular Longevity, 2016, 1404505.

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Microbial Growth and Viability Assay

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Nontypeable Haemophilus influenzae (Gram-negative, ATCC 49766), Moraxella catarrhalis (Gram-negative, ATCC 49143), Streptococcus pneumoniae (Gram-positive, ATCC 6301), and Pseudomonas aeruginosa (Gram-negative, ATCC 14203) were all purchased from American Type Culture Collection (ATCC) and stored in glycerol at –80°C. Brain heart infusion (BHI) media was purchased from Fisher Scientific and agar, hemin from bovine ≥ 90%, β-Nicotinamide adenine dinucleotide hydrate (NAD), and paraformaldehyde (PFA) were purchased from Sigma Aldrich. The Filmtracer™ LIVE/DEAD™ Biofilm Viability Kit was obtained from ThermoFisher Scientific. Phosphate buffer saline (1x) was prepared in distilled water and autoclaved.

Locke A.K., Zaki F.R., Fitzgerald S.T., Sudhir K., Monroy G.L., Choi H., Won J., Mahadevan-Jansen A, & Boppart S.A. (2022). Differentiation of otitis media-causing bacteria and biofilms via Raman spectroscopy and optical coherence tomography. Frontiers in Cellular and Infection Microbiology, 12, 869761.

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Propagation of Common Otitis Media Pathogens

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The three main bacteria that cause acute otitis media (AOM) were purchased from American Type Culture Collection (ATCC): nontypeable Haemophilus influenzae (ATCC #49766), Moraxella catarrhalis (ATCC #49143), and Streptococcus pneumoniae (ATCC #6301). Propagation methods as recommended by ATCC were used for each strain in preparation for bacteria cultures. Each bacterial species was streaked separately onto MH agar and chocolate agar plates. H. influenzae is a fastidious organism that requires lysed red blood cells not found in MH agar, therefore it is commonly grown on chocolate agar. To be effectively grown on MH agar, hemin and nicotinamide adenine dinucleotide (NAD)-rich disks (Hardy Diagnostics, Santa Maria, CA) were added to MH agar plates using steel tweezers that were disinfected between the additions of disks. Chocolate and MH agar plates were cultured for 24 hours at 37 °C with 5% CO₂.

Ayala O.D., Wakeman C.A., Pence I.J., O’Brien C.M., Werkhaven J.A., Skaar E.P, & Mahadevan-Jansen A. (2017). Characterization of bacteria causing acute otitis media using Raman microspectroscopy. Analytical methods : advancing methods and applications, 9(12), 1864-1871.

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Analytical Sensitivity and Cross-Reactivity Evaluation of AdvanSure Assay

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Serially diluted plasmids containing the target gene were used for determination of analytical sensitivity. pGEM^®-t easy vector (Promega, Madison, WI, USA) was used for plasmid DNA preparation. Serial dilutions of the prepared plasmid DNA were made from 10⁷ to 10⁰ copies per reaction to determine the analytical sensitivity of the assay. The plasmid DNAs of RSV A (Korea Bank for Pathogenic Viruses, KBPV-VR-41), PIV 1 (KBPV-VR-44), HRV (KBPV-VR-39), CoV 229E (KBPV-VR-9), CoV OC43 (KBPV-VR-8), and BoV (clinical isolate) were used. Three replicates of each dilution step were performed. The lower detection limit was defined as the lowest concentration detected by the assay in a series of three replicates.
The cross-reactivity of the AdvanSure assay was evaluated using nine different types of bacteria. Streptococcus pneumonia (ATCC 49619), Streptococcus pyogenes (ATCC 19615), Staphylococcus epidermidis (ATCC 12228), Moraxella catarrhalis (clinical isolate), Pseudomonas aeruginosa (ATCC 27853), Escherichia coli (ATCC 25922), Klebsiella pneumonia (ATCC 700603), Staphylococcus aureus (ATCC 25923), and Haemophilus influenza (ATCC 9007) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). The RNA or DNA of supplied samples was extracted and assayed with the AdvanSure assay abiding by the same procedures applied for sample processing.

Jung Y.J., Kwon H.J., Huh H.J., Ki C.S., Lee N.Y, & Kim J.W. (2015). Comparison of the AdvanSure™ real-time RT-PCR and Seeplex® RV12 ACE assay for the detection of respiratory viruses. Journal of Virological Methods, 224, 42-46.

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