Lyophilized pellets of Escherichia coli (ATCC 25922, and ATCC 700609) were purchased from ATCC. Tryptic Soy Broth (TSB), and Tryptic Soy Agar (TSA) were purchased from Sigma Aldrich; Nutrient Broth (NB) and Nutrient Agar (NA) were bought from Beckton Dickinson. Highly-purified glycerol was purchased from MP Biomedicals. Phosphate buffer saline (1X PBS) (without Calcium, Magnesium) was purchased from Lonza Walkersville Inc. Nalidixic Acid Sodium Salt was purchased from Alfa Aesar. Human urine (male, viral negative) was purchased from BioreclamationIVT.
Nalidixic acid sodium salt
Manufactured by Thermo Fisher Scientific
Sourced in United States
Nalidixic acid sodium salt is a synthetic antibacterial compound used in laboratory settings. It functions as a quinolone antibiotic that inhibits bacterial DNA gyrase, an essential enzyme for bacterial cell division and growth. This product is primarily used for research and scientific applications.
Automatically generated - may contain errors
3 protocols using nalidixic acid sodium salt
1
Bacterial Culture Preparation and Characterization
2
Nalidixic Acid-Resistant Salmonella Preparation
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Since similar findings were observed with both strains in the in vitro studies, S. Heidelberg 1904 was used for further investigations. To avoid potential competition from the skin and meat microflora, the strain was induced for resistance to nalidixic acid at 50 µl/mL nalidixic acid sodium salt (NA; CAS No. 3374-05-8, Alfa Aesar, Haverhill, MA) for selective enumeration as previously described (Nair and Kollanoor Johny, 2017b (link)). After the growth of culture to 9 log10 CFU/mL was confirmed by plating, an overnight culture was pelleted by centrifugation (3,600 x g for 15 min at 4°C) and suspended in sterile PBS to obtain a final concentration of 4 log10 CFU/50 µl.
3
Salmonella Heidelberg Isolation and Enumeration
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An MDR S. Heidelberg isolate from the 2011 ground turkey outbreak in the United States (kindly donated by Dr. Irene Hanning, College of Genome Sciences and Technology, University of Tennessee and Dr. Venkitanarayanan, University of Connecticut), was used for the study. Working cultures of S. Heidelberg was prepared from the glycerol stock cultures stored at -80°C. S. Heidelberg was made resistant to 50 μg/ml nalidixic acid sodium salt (NA; CAS. no. 3374-05-8, Alfa Aesar, Haverhill, MA, United States) for selective enumeration. The growth of S. Heidelberg (overnight culture) in tryptic soy broth (TSB; catalog no.C7141, Criterion, Hardy Diagnostics, Santa Maria, CA, United States) was determined on xylose lysine desoxycholate agar plates (XLD; catalog no. C 7322, Criterion, Hardy Diagnostics, Santa Maria, CA, United States) containing, 50 μg/ml NA and incubating at 37°C for 24 h. Then, final inoculum levels of 4 and 7 log10 CFU/ml were prepared from overnight broth culture (∼9 log10 CFU/ml) after centrifugation (3,600 × g, 15 min, 4°C) (Allegra X-14 R centrifuge, Beckman Coulter; 5350 Lakeview Parkway S Drive, Indianapolis, IN, United States) and suspending the pellets in sterile phosphate-buffered saline (PBS; pH 7.2) (Kollanoor-Johny et al., 2012a (link)).
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