Mice were given a cocktail of ciprofloxacin.HCl (TCI, Portland, Oregon, 0.15 g/liter), gentamycin sulfate (0.2 g/liter), bacitracin (1g/liter) and streptomycin (2 g/liter) (Thermo-Fisher, Waltham, MA) in drinking water ad libitum. In some experiments, niacin (10 mg/ml) was given in drinking water ad libitum for the indicated period of time.
Bacitracin
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom, United States
Bacitracin is a type of antibiotic that is commonly used in laboratory settings. It is a polypeptide antibiotic produced by the bacterium Bacillus licheniformis. Bacitracin functions by interfering with the synthesis of the bacterial cell wall, which can lead to cell death.
Automatically generated - may contain errors
Lab products found in correlation
Streptomycin, by Thermo Fisher Scientific (11 mentions)
Vancomycin, by Thermo Fisher Scientific (6 mentions)
Ampicillin, by Thermo Fisher Scientific (5 mentions)
Chloramphenicol, by Thermo Fisher Scientific (4 mentions)
Kanamycin, by Thermo Fisher Scientific (3 mentions)
Imipenem, by Thermo Fisher Scientific (3 mentions)
Penicillin g, by Thermo Fisher Scientific (3 mentions)
Nalidixic acid, by Thermo Fisher Scientific (3 mentions)
Rifampicin, by Thermo Fisher Scientific (2 mentions)
Ofloxacin, by Thermo Fisher Scientific (2 mentions)
Fosfomycin, by Thermo Fisher Scientific (2 mentions)
Gentamicin, by Thermo Fisher Scientific (2 mentions)
Amoxicillin, by Thermo Fisher Scientific (2 mentions)
Tetracycline, by Thermo Fisher Scientific (2 mentions)
Ciprofloxacin, by Thermo Fisher Scientific (2 mentions)
Clindamycin, by Thermo Fisher Scientific (2 mentions)
Polymyxin b, by Thermo Fisher Scientific (2 mentions)
Gentamycin sulfate, by Thermo Fisher Scientific (1 mentions)
Optochin disc, by Thermo Fisher Scientific (1 mentions)
Trypticase soy agar, by Thermo Fisher Scientific (1 mentions)
15 protocols using bacitracin
1
Antibiotic Cocktail for Mice
2
Broad-spectrum Antibiotic Treatment Protocol
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The antibiotics cocktail containing ciprofloxacin (TCI, Portland, Oregon, 0.15 g/liter), gentamicin (0.2 g/liter), bacitracin (1g/liter) and streptomycin (2 g/liter) (Thermo-Fisher, Waltham, MA) in drinking water was given to mice ad libitum.
3
Isolation of Las Bacteria from Psyllids, Citrus Seeds, and Dodder
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About 80 Las‐infected adult psyllids (Ct value = 26 to 32) were collected from the psyllid room (CREC‐UF) and used to dissect midguts in sterile PBS under a dissecting stereomicroscope by using depressed glass wells and fine entomological needles. After tissue samples were dissected, the tissues were washed three times with PBS and transferred to a sterile Eppendorf tube containing 0.3% bacitracin in PBS (Ghanim et al., 2016 ). Samples were pipetted up and down slowly to release the bacteria cells attached to the midgut.
Selected seeds isolated from infected and symptomatic citrus plants (Ct value about 27 to 29) were washed five times in sterile water, dried in filter paper and the coats were removed by using high‐precision straight fine tweezers. Seed coats were divided into very small pieces and suspended with 0.3% bacitracin (Thermo Fisher, Waltham, MA, USA) in PBS.
Individual dodder tendrils positive for Las (Ct = 24 to 26) were removed intact from parasitized plants. A segment of 10 mm was removed from the centre of dodder stem pieces, washed five times in sterile water, dried in paper filter, transferred to 0.3% bacitracin in PBS and cut into very small pieces with razor blades. The mixtures were pipetted up and down slowly to release the bacteria. Solution containing Las was transferred to a sterile Eppendorf tube.
Selected seeds isolated from infected and symptomatic citrus plants (Ct value about 27 to 29) were washed five times in sterile water, dried in filter paper and the coats were removed by using high‐precision straight fine tweezers. Seed coats were divided into very small pieces and suspended with 0.3% bacitracin (Thermo Fisher, Waltham, MA, USA) in PBS.
Individual dodder tendrils positive for Las (Ct = 24 to 26) were removed intact from parasitized plants. A segment of 10 mm was removed from the centre of dodder stem pieces, washed five times in sterile water, dried in paper filter, transferred to 0.3% bacitracin in PBS and cut into very small pieces with razor blades. The mixtures were pipetted up and down slowly to release the bacteria. Solution containing Las was transferred to a sterile Eppendorf tube.
4
Antibiotic Resistance Genes and Susceptibility
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Antibiotic resistance genes (ARGs) were predicted using the BLASTn method against the Comprehensive Antibiotic Resistance Database (CARD) [51 (link)] using an identity cut-off of 70% and an E-value < 1.0 E-6. Antibiotic susceptibility test was done using the Kirby-Bauer disk diffusion method on Mueller Hinton agar according to the Clinical and Laboratory Standards Institute (CLSI). Commercially prepared antibiotic disks used in this study were kanamycin (30 μg), streptomycin (10 μg), imipenem (10 μg), vancomycin (10 μg), ofloxacin (5 μg), ampicillin (30 μg), penicillin G (10 iu), rifampicin (5 μg), bacitracin (10 μg), fosfomycin (50 μg), and chloramphenicol (30 μg) (Thermo Fisher Scientific, MA, USA).
5
Microbial Identification in Sputum Samples
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Microbial identification by sputum culture, gram staining and standard biochemical tests was performed by medical laboratory technologists in accordance with the Clinical and Laboratory Standard Institute standards. Observation of haemolytic reaction, growth of gram negative and Haemophilus species bacteria were identified by culturing the good quality sputa (>25 leukocytes and <10 squamous epithelial cells/low power field) onto trypticase soy agar with 5% sheep blood, McConkey agar and chocolate agar with bacitracin (Thermo Scientific, Malaysia), respectively. The cultured media were incubated at 37 °C overnight in the presence of 5% carbon dioxide. Further confirmation tests include biochemical tests for gram negative bacteria, optochin discs (Oxoid, United Kingdom) for Streptococcus pneumoniae, X (hemin), V (nicotinamide adenine dinucleotide) and X + V factor discs (Oxoid, United Kingdom) for Haemophilus species differentiation, latex agglutination test for Staphylococcus aureus, and Vitek II system (BioMérieux, France) to further validate the organism. Simultaneously, the presence of Mycobacterium tuberculosis in the sputum specimens was also checked by using Ziehl–Neelsen staining, followed by cultivation on Ogawa medium for at least two weeks at 37 °C.
6
Antibiotic Resistance Profiling
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Antibiotic resistance genes (ARGs) were predicted using the BLASTn method against the Comprehensive Antibiotic Resistance Database (CARD) [49] using an identity cut-off of 70%
and an E-value < 1.0 E -6 . Antibiotic susceptibility test was done using the Kirby-Bauer disk diffusion method on Mueller Hinton agar according to the Clinical and Laboratory Standards Institute (CLSI). Commercially prepared antibiotic disks used in this study were kanamycin (30 μg), streptomycin (10 μg), imipenem (10 μg), vancomycin (10 μg), ofloxacin (5 μg), ampicillin (30 μg), penicillin G (10 iu), rifampicin (5 μg), bacitracin (10 μg), fosfomycin (50 μg), and chloramphenicol (30 μg) (Thermo Fisher Scientific, MA, USA).
and an E-value < 1.0 E -6 . Antibiotic susceptibility test was done using the Kirby-Bauer disk diffusion method on Mueller Hinton agar according to the Clinical and Laboratory Standards Institute (CLSI). Commercially prepared antibiotic disks used in this study were kanamycin (30 μg), streptomycin (10 μg), imipenem (10 μg), vancomycin (10 μg), ofloxacin (5 μg), ampicillin (30 μg), penicillin G (10 iu), rifampicin (5 μg), bacitracin (10 μg), fosfomycin (50 μg), and chloramphenicol (30 μg) (Thermo Fisher Scientific, MA, USA).
7
Antibiotic Susceptibility Assay of Lactic Acid Bacteria
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The antibiotics used for susceptibility assay were ampicillin (10 μg), clindamycin (2 μg), erythromycin (15 μg), gentamicin (10 μg), streptomycin (25 μg), tetracycline (30 μg), chloramphenicol (30 μg), kanamycin (30 μg), sulphamethoxazole/trimethoprim (25 μg), vancomycin (30 μg), ciprofloxacin (5 μg), amoxicillin (10 μg), bacitracin (10 μg), nalidixic acid (30 μg), and penicillin (10 μg) (Oxoid Ltd, England). The antibiotics were selected due to their common use in local animal farming. A total of 1 ml LAB culture grown in MRS broth was collected by centrifugation at 1000 × g for 5 min. The cell pellet was collected and washed twice using 1 ml of 0.85% (w/v) NaCl, followed by suspending the cell pellet with 0.5 ml of 0.85% (w/v) NaCl. The cell suspension was adjusted to 0.5 Mc Farland by using 2 ml of NaCl 0.85% (w/v) prior to spread plate on MRS agar. The antibiotic disc was then placed on MRS agar plate. The diameter of inhibitory zones was measured after 48 h of incubation at 30°C under anaerobic condition. The assay was conducted in triplicates [20 (link)].
8
Antimicrobial Susceptibility of Clinical Isolates
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Silver nitrate (AgNO3) was obtained from Saudi Overseas Marketing and Trading Company (SOMATCO), Riyadh, Saudi Arabia. Nutrient agar plates and nutrient broth (Difco, Becton, Dickinson and Company, Sparks Glencoe, MD, United States) as well as potato-dextrose agar (PDA) plates (Difco, Becton, Dickinson and Company, Sparks Glencoe, MD, United States) were purchased from Wateen Alhaiah Company, Riyadh, Saudi Arabia. All clinical bacterial isolates were obtained from the Microbiology Laboratory of Princess Nourah bint Abdulrahman University, Riyadh, Saudi Arabia. Antibiotic discs were obtained from OXOID TM United Kingdom at the following concentrations: bacitracin, 10 μg/mL; ciprofloxacin, 10 μg/mL; tetracycline, 30 μg/mL; and cefixime, 5 μg/mL.
9
Antimicrobial Evaluation of Prosopis juliflora Extracts
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The disk diffusion method was used to characterize the antimicrobial effect of aqueous and ethanolic extracts of P. juliflora leaves and fruits on bacterial (E. coli, S. aureus, B. subtilis, and P. microbilis) and yeast (C. albicans) isolates. A microbial lawn of each isolate was spread on Mueller–Hinton agar (HiMedia-India). Sterile paper disks (Thermo Scientific Oxoid Blank Antimicrobial Susceptibility Disks) of 5 mm diameter were inoculated with 20 µl of different concentrations of each extracts (1, 5, 10, 20, and 50 mg/ml). Dry disks were then transferred with sterile forceps into the inoculated agar plates. Disks with sterile distilled water were used as negative controls and antibiotic disks were used as positive controls. The antibiogram of the bacterial isolates were also determined to compare the effect of PJ-WS-LE extract to commonly used antibiotics (Ampicillin (AMP), Amoxicillin (AMX), Bacitracin (B), Carbenicillin (CB), and Cephalothin (CR))(Oxoid-USA). Plates were incubated at 37 °C for 24 h. The experiment was conducted in triplicates44 (link).
10
Antibiotic Susceptibility Testing of Gonococci
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Kirby–Bauer disk diffusion susceptibility test protocol (Hudzicki, 2009 ) was adapted to test the toxicity of different chemical compounds for gonococci. Briefly, overnight cultures of N. gonorrhoeae were collected in 3 ml of sterile saline. OD was adjusted to 0.5 in the MacFarland scale and cells were plated (by streaking the swab three times over the entire agar surface) on GC with Kellogg and hemoglobin. Disks with antibiotics or other substances were then applied and plates incubated for 24 h at 37°C and 5% CO2. Then, the diameter of the inhibition zone of growth was measured in cm. Oxoid antibiotic susceptibility disks were used to test recombinant mutants and wt N. gonorrhoeae strain sensitivity to antibiotics. We used disks with miniscule 30 μg/ml, imipenem 10 μg/ml, bacitracin 10 μg/ml, azithromycin 15 μg/ml, gentamycin 30 μg/ml, and polymyxin B 300 μg/ml (Oxoid, United Kingdom).
We also used Whatman filter paper disks on which we applied 5 μl of 30% H2O2, Triton-X100, or 10% SDS. Tests were carried out at least three times and the results were averaged. The significance of results was determined as described in Section “Statistical analysis.”
We also used Whatman filter paper disks on which we applied 5 μl of 30% H2O2, Triton-X100, or 10% SDS. Tests were carried out at least three times and the results were averaged. The significance of results was determined as described in Section “Statistical analysis.”
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