Nutrient broth

Manufactured by Thermo Fisher Scientific

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Nutrient broth is a general-purpose culture medium used for the growth and cultivation of a wide variety of microorganisms, including bacteria, yeast, and fungi. It provides essential nutrients and growth factors necessary for the proliferation of microbial cells.

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Nutrient broth No. 2 (31 citations) Sterile nutrient broth (9 citations) Nutrient Broth medium (8 citations) Oxoid Nutrient Broth (4 citations) Nutrient Broth media (4 citations) Bacto nutrient broth (2 citations) Nutrient broth powder (2 citations) Nutrient broth nº 2 (2 citations) Oxoid Nutrient Broth No. 2 (2 citations) Nutrient broth; CM0001 (2 citations)

248 protocols using nutrient broth

Bacterial Survival on Vegetables

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Listeria monocytogenes and Staphylococcus aureus were used in this research. The strains belong to the Culture Collection of the Laboratory of Predictive Microbiology, Dept. SAFE, University of Foggia. They are wild isolates found in foods; some preliminary experiments showed that they could survive on vegetables.
The bacteria were stored at −20°C in Nutrient broth (Oxoid, Milan, Italy), supplemented with 33% of sterile glycerol; before each assay, they were grown in Nutrient broth, incubated at 37°C for 24 h. The microorganisms were centrifuged at 3,000 g for 10 min and the pellet was suspended in a brine prepared with tap water and 4% NaCl. The viable count of these suspensions was 7 log cfu/ml.

Bevilacqua A., Campaniello D., Speranza B., Sinigaglia M, & Corbo M.R. (2018). Survival of Listeria monocytogenes and Staphylococcus aureus in Synthetic Brines. Studying the Effects of Salt, Temperature and Sugar through the Approach of the Design of Experiments. Frontiers in Microbiology, 9, 240.

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Bacillus Strains from Stingless Bee Honey

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A total of 23 Bacillus strains isolated from stingless bee honey were included in this study. Pathogenic strains (Escherichia coli, Salmonella thyphimurium, Klebsiella pneumonia, Pseudomonas aeruginosa, and Staphylococcus aureus) were kindly supplied by the Enzyme and Microbial Technology Research Center, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Malaysia. The strains were maintained at −80 °C in nutrient broth (Oxoid, Basingstoke, UK) with 20% (v/v) glycerol and were propagated three times in nutrient broth for activation prior to experimental use.

Zulkhairi Amin F.A., Sabri S., Ismail M., Chan K.W., Ismail N., Mohd Esa N., Mohd Lila M.A, & Zawawi N. (2019). Probiotic Properties of Bacillus Strains Isolated from Stingless Bee (Heterotrigona itama) Honey Collected across Malaysia. International Journal of Environmental Research and Public Health, 17(1), 278.

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Bacterial Zinc Resistance Cultivation

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Highly zinc-resistant bacterial strains were grown in nutrient broth (Oxoid, UK) with .1% zinc salt (ZnSO₄.7H₂O) and without zinc used as controls. nutrient broth (Oxoid, UK) was prepared and supplemented with zinc, autoclaved at 121°C, 15-pound pressure for 20 minutes (Samheung, Korea). Bacterial colonies were inoculated into the sterilized nutrient broth on the flasks. The flasks were kept at 30°C for 24 hours at 150 rpm, shaking in a rotator shaker (Thermo Scientific™, UK). After 24 hours, flasks were taken out of the shaker and centrifuged test and controlled at 10 000 g (Thermo Scientific™, UK) for 10 minutes. Discarded the supernatant and stored the pellet for FT-IR.

Yasmin R., Zafar M.S., Tahir I.M., Asif R., Asghar S, & Raza S.K. (2022). Biosorptive Potential of Pseudomonas species RY12 Toward Zinc Heavy Metal in Agriculture Soil Irrigated with Contaminated Waste Water. Dose-Response, 20(3), 15593258221117352.

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Modified Carbapenem Inactivation Assay for Carbapenemase Detection

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Isolates that were resistant and intermediate to either meropenem or imipenem were tested for production of carbapenemase by using the modified carbapenem inactivation method (CIM) as described by CLSI;26 . Briefly; 1µL loop full colony of test isolate from overnight blood agar plate was suspended in 2 mL of nutrient broth (Oxoid UK) and 10µg of meropenem disk was added to the nutrient broth and fully immersed. The tubes were incubated at 37°C in ambient air without agitation for 4 h ± 15 min. Subsequently, the meropenem disks were removed using a 10 µL inoculation loop and applied to Mueller-Hinton agar plates (Oxoid, UK) freshly inoculated with a 0.5 McFarland suspension of a carbapenem-susceptible strain (Escherichia coli ATCC^® 25922) after overnight incubation results were interpreted as described by CLSI.26 ,27 (link)

Seman A., Mihret A., Sebre S., Awoke T., Yeshitela B., Yitayew B., Aseffa A., Asrat D, & Abebe T. (2022). Prevalence and Molecular Characterization of Extended Spectrum β-Lactamase and Carbapenemase-Producing Enterobacteriaceae Isolates from Bloodstream Infection Suspected Patients in Addis Ababa, Ethiopia. Infection and Drug Resistance, 15, 1367-1382.

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Acid-Adapted Listeria monocytogenes Strain

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Listeria monocytogenes 56 LY used as the target strain in this study was provided by the Food Microbiology Laboratory of the University of Bologna (Cesena, Italy). It was firstly sub-cultured thrice in Nutrient Broth (CM0001, Oxoid Ltd., Basingstoke, UK) at 37 °C for 24 h. This was followed by an acid-adaptation through a new culture in Nutrient Broth adjusted at pH 6.5 using citric acid. This specific condition was observed in a previous study to strongly enhance the mild thermal tolerance of this strain (Tchuenchieu et al., 2018c (link)).

Tchuenchieu A., Sado Kamdem S., Bevivino A., Etoa F.X, & Essia Ngang J.J. (2022). Development of a predictive model of the microbial inactivation of L. monocytogenes during low thermal treatment of fruit juices in combination with carvacrol as aroma compound. Current Research in Food Science, 5, 374-381.

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Isolation and Identification of Bacterial Pathogens from Diseased African Catfish

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The bacteria used for the antimicrobial analyses were obtained from samples (gills, kidneys and skin) collected from diseased African cat fish from various farms in Kwara state, Nigeria. The samples were analyzed at the Veterinary Microbiology laboratory, University of Ilorin, Nigeria according to method of Fagbemi et al. (2009) . Briefly, 1 g of each sample was pre-enriched in 9 ml of buffered peptone water (Oxoid, Hampshire, UK) incubated at 37 0 C for 24 hours. Pre-enriched sample (1ml) was then inoculated into 9 ml of nutrient broth (Oxoid, Hampshire, UK) and incubated overnight at 37 0 C. The turbid nutrient broth was subsequently inoculated unto 5 % sheep blood agar (Oxoid, Hampshire, UK) incubated aerobically at 37 0 C for another 24 hours. The resultant colonies were purified on nutrient agar (Oxoid, Hampshire, UK) and then subjected to Gram staining and biochemical characterization including oxidase, catalase and coagulase tests (Cowan and Steel, 2002) . The isolates were kept in 20 % glycerol broth at -20 0 C for future use.

Ajadi A., Emikpe B.O., & Ahmed A.O. (2021). Invitro Antimicrobial Activities of Mitracarpus scaber Against Some Common Bacteria of Aquatic Origin. Media Kedokteran Hewan, 32(3), 119-119.

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Isolation and Identification of Staphylococci

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The clinical samples were inoculated into freshly prepared nutrient broth (Oxoid Ltd., Basingstoke, Hampshire, England) and incubated at 37 °C for 24 h. A loop-full of growth from the nutrient broth medium was streaked onto Mannitol Salt Agar (Oxoid Ltd., Basingstoke, Hampshire, England) and incubated for 24 h at 37 °C (Public Health England, 2017). Discrete, round, single, goldenyellow colonies were presumptively identified as Staphylococci. The isolates were characterized by conducting the following tests: gram stain, catalase, coagulase (Ochei and Kolhatkar, 2008; Chandra and Mani, 2011) . Microgen™ Staph-ID System was used to identify the isolates.

Oche D.A., Abdulrahim U., Oheagbulem A., & Olayinka B.O. (2021). Isolation of Biofilm Producing Methicillin-Resistant Staphylococcus aureus from Hospitalized Orthopaedic Patients in Kano State, Nigeria. Nigerian Journal of Basic and Applied Sciences, 28(1), 66-74.

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Isolation and Characterization of S. aureus from CRS

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Investigation of S. aureus strains isolated from CRS patients was approved by The Queen Elizabeth Hospital Human Ethics Committee. Written informed consent was obtained for all study participants. To isolate S. aureus from sinonasal swabs, Columbian blood agar plates, colistin nalidixic acid plates or cystine lactose electrolyte deficient plates (all from Thermofisher Scientific, Australia) were employed. Latex agglutination tests and antibiotic sensitivity testing were performed by the commercial laboratory Adelaide Pathology Partners and used to confirm identification of S. aureus and MRSA strains (data not shown). Antibiotic resistance was determined using disc diffusion methods as according to Clinical and Laboratory Standards Institute (CLSI) recommendations (CLSI, 2012 ). Isolates were then subcultured in nutrient broth overnight (Thermo Fisher, Scoresby, Victoria, Australia) and stored in nutrient broth with 20% glycerol at −80°C.

Drilling A.J., Ooi M.L., Miljkovic D., James C., Speck P., Vreugde S., Clark J, & Wormald P.J. (2017). Long-Term Safety of Topical Bacteriophage Application to the Frontal Sinus Region. Frontiers in Cellular and Infection Microbiology, 7, 49.

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Comprehensive Phytochemical Analysis

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All chemicals used were of analytical grade. Ethanol, Methanol, (±)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox), 2,2-Diphenyl-1-picrylhydrazyl (DPPH), 2,4,6-tris(2-pyridyl)-s-triazine (TPTZ), Acetone, Aluminium Chloride, Ammonium Acetate, Catechin, Copper (II) Chloride Dehydrate, Ferric Chloride, Folin-Ciocalteur Reagent, Gallic Acid, Glacial Acetic Acid, Hexane, Hydrochloric Acid, Neocuprine, Potassium Hexacyanoferrate (II), Sodium Acetate Trihydrate, Sodium Carbonate, Sodium Hydrogen Sulphite, Sodium Hydroxide, Sodium Nitrate, and Zinc Acetate were purchased from Sigma-Aldrich (Castle Hill, Sydney, Australia). Nutrient Agar Plates and Nutrient Broth were purchased from Thermo Fisher Scientific (Thebarton, South Australia, Australia). Deionised (DI) water was collected following purification (Millipore Australia, North Ryde, NSW), with a resistivity higher than 18 MΩcm⁻¹.

Hunter M., Ghildyal R., D'Cunha N.M., Gouws C., Georgousopoulou E.N, & Naumovski N. (2021). The bioactive, antioxidant, antibacterial, and physicochemical properties of a range of commercially available Australian honeys. Current Research in Food Science, 4, 532-542.

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Interspecies Conjugation Protocol for E. coli and A. mediterranei

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All strains used in this study are listed in Table 1. E. coli strains were grown at 37 °C in LB. When required, antibiotics were added to E. coli cultures in liquid (or on solid) medium at the following concentrations: ampicillin, 50 µg/mL (or 100 µg/mL in solid); apramycin, 25 µg/mL (or 50 µg/mL); hygromycin, 50 µg/mL (or 150 µg/mL); kanamycin, 25 µg/mL in liquid or solid medium. A. mediterranei strains were grown at 30 °C on GYM agar medium (32) for sporulation before the preparation of spore stocks, in TSB (Tryptic Soy Broth, Becton Dickinson) for DNA extraction and in MP5 (33) for rifamycin production. Conjugations between E. coli ET12567 harboring pUZ8002 (or pUZ8003) and A. mediterranei were carried out according to Kieser et al. (34) using MS medium complemented with 10 mM CaCl2 (35), instead of MgCl2. An overlay (3 mL) of soft nutrient agar (Nutrient Broth (Thermo Scientific, Dardilly, France), with 0.8% agar) containing 25 µg/mL nalidixic acid and the appropriate antibiotics for the selection of exconjugants was added after overnight incubation of the conjugation plates. The plates were incubated for 5–7 days until exconjugants clones became visible. Antibiotic concentration used for A. mediterranei were as follows: apramycin, 50 µg/mL; erythromycin, 75 µg/mL; hygromycin, 75 µg/mL.

Santos L.D., Caraty-Philippe L., Darbon E, & Pernodet J.L. (2022). Marker-Free Genome Engineering in Amycolatopsis Using the pSAM2 Site-Specific Recombination System. Microorganisms, 10(4), 828.

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