Sterile nutrient broth

Manufactured by Thermo Fisher Scientific

Sourced in United Kingdom

Sterile nutrient broth is a liquid growth medium used for the cultivation of a wide range of microorganisms. It provides essential nutrients to support the growth and proliferation of various bacterial, fungal, and other microbial species.

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Nutrient broth (248 citations)

9 protocols using sterile nutrient broth

Quality Assurance of Decellularized Tendons

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Decellularised split pSFT were assessed for sterility, removal of cellular material, maintenance of histological structure, total DNA content, biocompatibility and biomechanical properties.
Sterility of oSFT was tested immediately following extraction using a small piece of tissue from both the toe and ankle regions. For decellularised split pSFT, one half of the tendon was designated for QA and the other for potential use in the animal study. Sterility of pSFT was assessed following decellularisation and prior to packaging for every tendon produced. This assessment was carried out by placing a small sample of the toe region of each QA tendon in 10 ml sterile nutrient broth (Oxoid), incubated at 37 °C for 48 h and checked for signs of cloudiness or film formation. Only tendons where the QA sample showed no signs of microbial growth were used in the animal study. For all other quality assurance analyses, tissue was taken from a set of 6 QA tendon halves, which were chosen at random from the packaged tissues. As the decellularisation process had been previously established [26 (link)], it was deemed sufficient to perform checks to confirm decellularisation on a random subset of the tendons.

Edwards J.H., Jones G.L., Herbert A., Fisher J, & Ingham E. (2021). Integration and functional performance of a decellularised porcine superflexor tendon graft in an ovine model of anterior cruciate ligament reconstruction. Biomaterials, 279, 121204.

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Standard Bacterial Strain Cultivation

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Standard strains, Escherichia coli ATCC (American Type Culture Collection) 25922, Salmonella Enteritidis ATCC 13076, Listeria monocytogenes ATCC 19114 were tested as controls. They were grown in a test tube containing 10 mL sterile nutrient broth (Oxoid Ltd., Basingstoke, Hampshire, England) at 37 °C for 24 h. A loopful of inoculum was transferred onto selective media: TBX agar for E. coli, XLD agar for Salmonella Enteritidis (Oxoid Ltd., Basingstoke, Hampshire, England) and Palcam agar base supplemented with Listeria Palcam antimicrobic supplement (Oxoid Ltd., Basingstoke, Hampshire, England) for Listeria monocytogenes. Plates were incubated for 24 h at 37 °C. Bacterial morphology was confirmed by optical microscopy.

Cerezo A.B., Cătunescu G.M., González M.M., Hornedo-Ortega R., Pop C.R., Rusu C.C., Chirilă F., Rotar A.M., Garcia-Parrilla M.C, & Troncoso A.M. (2020). Anthocyanins in Blueberries Grown in Hot Climate Exert Strong Antioxidant Activity and May Be Effective against Urinary Tract Bacteria. Antioxidants, 9(6), 478.

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Antimicrobial Efficacy Testing of CREC and MRSA

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Target organisms: Clinical isolates of carbapenem-resistant E. coli (CREC) and methicillin-resistant Staphylococcus aureus (MRSA; EMRSA-15 variant B1). Preparation of bacteria: 10 µL of test bacteria was transferred with a loop to sterile nutrient broth (10 mL; Oxoid, UK), mixed thoroughly and incubated aerobically at 37 °C for ∼24 h (±4 h). Broth cultures were centrifuged at 3000 rpm (1500×g; Jouan CR3i centrifuge: Thermo, UK) for 10 min and the remaining pellet re-suspended in 10 mL of sterile distilled water (DW). During time-kill assays, the PAA antimicrobial agent was inactivated after the appropriate contact time by the addition of a neutralising solution [3% (w/v) Tween 80, 0.3% (w/v) Lecithin, 1.0% (w/v) Sodium thiosulfate, 1.5% (w/v) K₂HPO₄, KH₂PO₄ 0.05% (w/v), 1% (w/v) Poly-[sodium-4-styrenesulfonate], 0.1% (v/v) Triton® X100 (Sigma-Aldrich, UK) and prepared in phosphate-buffered saline (PBS) solution (Oxoid, UK)]. Neutralising solutions were sterilised by autoclaving (121 °C for 15 min) and refrigerated (2–8 °C) until required. All microbiology studies were conducted by the Environmental Research Laboratory, University College Hospital.

Sofokleous P., Ali S., Wilson P., Buanz A., Gaisford S., Mistry D., Fellows A, & Day R.M. (2017). Sustained antimicrobial activity and reduced toxicity of oxidative biocides through biodegradable microparticles. Acta Biomaterialia, 64, 301-312.

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Microbial Cultivation in Nutrient Broth

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One single colony of each type of microorganism (from the nutrient agar stock culture) was inoculated with a sterile loop, and is transferred into 10 mL sterile nutrient broth (Oxoid). The broth cultures were incubated in a shaking incubator at 37 °C for 16–20 h.

Chew Y.L., Mahadi A.M., Wong K.M, & Goh J.K. (2018). Anti-methicillin-resistance Staphylococcus aureus (MRSA) compounds from Bauhinia kockiana Korth. And their mechanism of antibacterial activity. BMC Complementary and Alternative Medicine, 18, 70.

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Microbial Cultivation and Enumeration

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The following microorganisms were tested: Staphylococcus aureus ATCC 6538P, Escherichia coli ATCC 25922, Salmonella enteritidis ATCC 13076 and Listeria monocytogenes ATCC 19114. Each strain was grown in a test tube containing 10 mL sterile nutrient broth (Oxoid Ltd., Basingstoke, Hampshire, UK) at 37 °C for 24 h. The purity of the inoculum was confirmed by microscopic examination of the Gram-stained smear. A loopful of inoculum was transferred to selective medium: Baird-Parker agar base supplemented with egg yolk tellurite emulsion for S. aureus, TBX agar for E. coli, XLD agar for Salmonella enteritidis (Oxoid Ltd., Basingstoke, Hampshire, UK) and Palcam agar (Oxoid Ltd., Basingstoke, Hampshire, UK) for Listeria monocytogenes. Plates were incubated for 24 h at 37 °C. Bacterial morphology was confirmed by optical microscopy. Several colonies were transferred to sterile saline solution (8.5 g/L), and adjusted to match the turbidity of McFarland 0.5 standard (1.5 × 10⁸ CFU/mL) [36 (link),37 (link)].

Vârban D., Zăhan M., Pop C.R., Socaci S., Ștefan R., Crișan I., Bota L.E., Miclea I., Muscă A.S., Deac A.M, & Vârban R. (2022). Physicochemical Characterization and Prospecting Biological Activity of Some Authentic Transylvanian Essential Oils: Lavender, Sage and Basil. Metabolites, 12(10), 962.

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Antibiotic Susceptibility of Enterococci

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Antimicrobial susceptibility testing for each isolate was performed using the Kirby–Bauer disk diffusion method on Muller–Hinton agar (OXOID, UK). From pure culture, selected colonies of bacteria were taken and transferred to a tube containing sterile nutrient broth (OXOID, UK) and incubated at 37°C until the turbidity of the suspension becomes adjusted to a McFarland standard (Hardy diagnostics, Canada) 0.5. Then, the suspension was spread evenly on Muller–Hinton agar based on Clinical and Laboratory Standards Institute (CLIS). The medium after exposed to a concentration gradient of antibiotic discs were incubated at 37°C for 18–24 hours. Grades of susceptibility pattern were recognized as sensitive and resistant by comparison of zone of inhibition according to the CLSI guidelines. Antimicrobial susceptibility patterns of Enterococci were assessed against the following antibiotic discs: vancomycin (30 μg), ampicillin (10 μg), nitrofurantoin (300 μg), tetracycline (30 μg), ciprofloxacin (5 μg), chloramphenicol (30 μg), erythromycin (15 μg), norfloxacin (30 μg), and doxycycline (30 μg) (all from Abtek Biologicals Ltd.).

Agegne M., Abera B., Derbie A., Yismaw G, & Shiferaw M.B. (2018). Magnitude of Vancomycin-Resistant Enterococci (VRE) Colonization among HIV-Infected Patients Attending ART Clinic in West Amhara Government Hospitals. International Journal of Microbiology, 2018, 7510157.

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Prevalence of Staphylococcus aureus in Clinical Samples

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A total of one hundred and fifty-four (154) clinical samples were collected from fifteen (15) hospitals in and around Nsukka, Southeastern Nigeria, between March and July 2021. Of the 154 clinical samples, 49 (36.84%), 103 (31.58%), and 3 (21.05%) were from wound/pus, urine, and ear swabs, respectively. The study was done per the Declaration of Helsinki [13 (link)], and appropriate approval (FPSRE/UNN/20/0008) was sought from the Research Ethics Committee of the Faculty of Pharmaceutical Sciences, University of Nigeria, Nsukka. Informed consent was also obtained from the patients before sample collection. Wound/pus swabs and ear swabs were inoculated into the sterile nutrient broth (Oxoid, UK) and incubated for 24 h at 37 ^oC, after which a loopful of the broth culture of each sample was inoculated onto sterile mannitol salt agar (Oxoid, UK) and incubated at 37 ^oC for 24 h. Urine samples were inoculated directly onto sterile mannitol salt agar plates. After incubation, colonies with yellowish pigments from the agar were characterized using standard microbiological techniques. The presumptive S. aureus isolates were subjected to the polymerase chain reaction (PCR) targeting the S. aureus-specific nuc gene [14 (link)].

Agbo M.C., Ezeonu I.M., Onodagu B.O., Ezeh C.C., Ozioko C.A, & Emencheta S.C. (2024). Antimicrobial resistance markers distribution in Staphylococcus aureus from Nsukka, Nigeria. BMC Infectious Diseases, 24, 320.

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Antimicrobial Efficacy of CuNP Foams

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Escherichia coli ATCC 25922, Staphylococcus aureus FDA 209P (MSSA, with methicillin resistance 0.125 µg/mL), and Kluyveromyces marxianus CBS 608 were selected as target microorganisms. Aliquots of the different lyophilized microorganisms were reconstituted in 0.9% NaCl solution, added to 20 mL of sterile nutrient broth (Oxoid) and incubated for 24 h at the optimal temperature for microbial growth, which was 37 °C for Staphylococcus aureus, 42 °C for Escherichia coli, and 28 °C for Kluyveromyces marxianus. Each sample was then diluted 10⁵ times by saline solution (pH = 6.4, [Cl⁻] = 0.15 M). 5 mL of each culture broth were left in contact with each type of CuNP treated foams, and incubated for 24 h, at the respective optimal growth temperatures. Untreated samples were used as controls. After the selected incubation times, 1 mL of culture broth was taken from each vessel and inoculated in a Petri dish containing nutrient agar. After 24 h, bacterial colony count was performed on each Petri dish, in order to assess the entity of bacterial growth inhibition exerted by treated polyurethane foams.

Sportelli M.C., Picca R.A., Ronco R., Bonerba E., Tantillo G., Pollini M., Sannino A., Valentini A., Cataldi T.R, & Cioffi N. (2016). Investigation of Industrial Polyurethane Foams Modified with Antimicrobial Copper Nanoparticles. Materials, 9(7), 544.

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Antimicrobial Susceptibility Testing of Enterococci

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Each isolate’s antimicrobial susceptibility was tested using the Kirby-Bauer disk diffusion method on Muller-Hinton agar (Oxoid Ltd, UK). Well-isolated pure colonies of the same morphological type were selected from the Bile Azide Agar plate and preserved on Tryptone soya broth (Sisco Research Laboratories Pvt., India). Selected bacterial colonies were taken from pure cultures and put into a tube with sterile nutrient broth (Oxoid Ltd, UK). The tube was then incubated at 37°C until the suspension’s turbidity was adjusted to a McFarland standard (Hardy diagnostics, Canada) of 0.5. Then, the suspension was spread evenly on Muller–Hinton agar. The medium was incubated at 37°C for between 18 and 24 hours after being exposed to an antibiotic concentration gradient on discs. The following antibiotic discs were used to test the antimicrobial susceptibility of enterococci: vancomycin (30 µg), ampicillin (10 µg), penicillin (10 units), doxycycline (30 µg), chloramphenicol (30 µg), erythromycin (15 µg),12 (link) erythromycin (5 µg), tetracycline (30 µg), ciprofloxacin (5 µg) and nitrofurantoin (300 μg).26 The status of resistance of the strain was determined by measuring the zone diameter (mm) and referring to CLSI standard for each drug of interest.27 ,28 (link)

Zike M., Ahmed A.M., Hailu A, & Hussien B. (2024). Vancomycin Resistant Enterococci Prevalence, Antibiotic Susceptibility Patterns and Colonization Risk Factors Among HIV-Positive Patients in Health-Care Facilities in Debre Berhan Town, Ethiopia. Infection and Drug Resistance, 17, 17-29.

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