Twenty-five grams of meat samples was homogenized for 90 s in a stomacher (Heidolph, Schwabach, Germany) with 225 ml of peptone water (PW) containing 6.5% NaCl and then incubated at 37°C for 24 h. After primary enrichment, a loopful (without shaking the flask) from each of the enriched homogenates was streaked onto Baird-Parker agar (MERCK, Darmstadt, Germany) supplemented with 5% egg yolk and tellurite and incubated under aerobic conditions at 37 °C for 24 h. Colonies with typical gray-black appearance surrounded by a clear zone were enumerated as coagulase-positive staphylococci and sub-cultured onto Mannitol salt agar. Clinical specimens were also cultured onto Brain Heart Infusion agar and Mannitol salt agar (MERCK, Darmstadt, Germany) [22 (link), 23 (link)]. The isolates were identified as S. aureus by further biochemical characterization using Gram stain, catalase, coagulase, oxidase, lipase, DNase, and PCR targeting the S. aureus-specific femA gene (S. aureus species specific).
Mannitol salt agar
Manufactured by Merck Group
Sourced in Germany, United States
Mannitol salt agar is a selective and differential culture medium used for the isolation and identification of Staphylococcus species, particularly Staphylococcus aureus, from clinical and non-clinical samples. It contains mannitol and sodium chloride, which inhibit the growth of most other bacteria, allowing Staphylococcus species to grow selectively.
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Lab products found in correlation
Blood agar, by Merck Group (11 mentions)
Mueller hinton broth (mhb), by Merck Group (7 mentions)
Mueller hinton agar (mha), by Merck Group (7 mentions)
Macconkey agar, by Merck Group (6 mentions)
Baird parker agar, by Merck Group (6 mentions)
Sabouraud dextrose agar, by Merck Group (5 mentions)
Buffered peptone water (bpw), by Merck Group (4 mentions)
Plate count agar, by Merck Group (3 mentions)
Gallic acid, by Merck Group (3 mentions)
Quercetin, by Merck Group (3 mentions)
Egg yolk tellurite emulsion, by Merck Group (3 mentions)
Sheep blood agar, by Merck Group (3 mentions)
Crystal violet, by Merck Group (3 mentions)
Agarose, by Merck Group (3 mentions)
Dna agar, by Merck Group (3 mentions)
Trypticase soy broth tsb, by Merck Group (3 mentions)
Eosin methylene blue emb, by Merck Group (2 mentions)
Nutrient broth, by Merck Group (2 mentions)
Sabouraud dextrose broth (sdb), by Merck Group (2 mentions)
Linoleic acid, by Merck Group (2 mentions)
53 protocols using mannitol salt agar
1
Staphylococcus aureus Isolation and Identification
2
Nasal Staphylococcus aureus Carriage in Healthy Ruminants
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During the period between January 2011 and May 2012, a total of 201 nasal swab samples were taken from healthy ruminants including 79 cattle, 78 sheep and 44 goats housed in three different areas (Khorramabad, Noor Abad, and Urmia) of Iran. For all animals, cotton swabs were inserted into the anterior nares of both left and right nostrils and were softly rolled against the inner walls. The Urmia University of Veterinary Ethical Committee approved the animal protocol for this study (K2-427). One swab was used to take a sample from both nostrils and then stored in Tryptone Soy Broth (TSB) (Merck, Germany) at 4°C prior to laboratory analysis. Cultures were incubated at 37°C for 24 hours and then streaked directly on plates of Mannitol Salt Agar (MSA) (Merck, Germany) and incubated at 37°C for 18 to 48 hours (23 ). The suspected S. aureus colonies were purified on sheep blood agar plates and then identified by standard biochemical tests including Gram staining, catalase reaction, oxidase reaction, mannitol fermentation and coagulase production (24 (link)). Staphylococcus aureus isolates were stored in TSB containing 15% glycerol at -20°C until needed.
3
Quantifying Bacterial Growth Inhibition
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The test bacterial suspension which has been standardized with McFarland 0.5 was diluted using 0.85% physiological NaCl. S. aureus or E. coli was grown in nutrient broth/NB (Merck) and then added 10 μL of yeast indigenous for 72 h at 30°C. 1 ml of bacterial suspension was platted in eosin methylene blue/EMB (Merck) for E. coli and mannitol salt agar/MSA (Merck) for S. aureus and then incubated at 37°C for 24 h. For positive (+) control treatment, 200 μL of bacterial culture of 1 × 104 CFU/mL + 2 μL amoxicillin 100 mg/mL were used, while 200 μL bacterial culture of 1 × 104 CFU/mL used as negative (-) control treatment. The population of E. coli and S. aureus were count every 24 h; all the treatments were replicated three times.
4
Bacterial Identification from Fly Samples
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Each fly was placed in 5 mL physiologic serum and was vortexed for 30 seconds. Then, 100 μl of physiologic serum was inoculated in the Brain Heart Infusion Agar culture medium. Inoculated Petri dishes were incubated for 24 hours at 37°C. After incubation, the appearance of the colonies was observed, and the number of colonies was counted.
One hundred μl physiologic serum was culture on the various culture media, i.e., Nutrient Agar, Brain Heart Infusion Agar, Cetrimide Agar, Brad Parker Agar (BPA) and MacConkey Agar (production of IBRESCO Company, Iran) and Mannitol Salt Agar (MSA) and Eosin Methylene Blue (EMB) (production of MERCK Company, Germany) to specify cultivation of P. aeruginosa, E. coli, and S. aureus. The various inoculated culture media were incubated for 24 hours at 37°C. After incubation, the number of colonies based on the specific characteristics of the bacteria was determined.
One hundred μl physiologic serum was culture on the various culture media, i.e., Nutrient Agar, Brain Heart Infusion Agar, Cetrimide Agar, Brad Parker Agar (BPA) and MacConkey Agar (production of IBRESCO Company, Iran) and Mannitol Salt Agar (MSA) and Eosin Methylene Blue (EMB) (production of MERCK Company, Germany) to specify cultivation of P. aeruginosa, E. coli, and S. aureus. The various inoculated culture media were incubated for 24 hours at 37°C. After incubation, the number of colonies based on the specific characteristics of the bacteria was determined.
5
Sterility Assessment of ADSC Culture
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Sterility of isolated cells was checked by harvesting 100 µl of conditioned medium from the flasks with ADSC culture and then its transfer to Petri dishes with various solid microbiological media: Blood Agar, Mannitol Salt Agar, MacConkey Agar, Müller-Hinton Agar, Anaerobic Blood Agar (all from Sigma-Aldrich Chemicals, Warsaw, Poland). Petri dishes were then incubated at 37 °C for 3 d to detect the growth of both aerobic and anaerobic microorganisms in medium samples.
6
Phytochemical and Antimicrobial Evaluation
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The L. perfoliatum seeds and chicory were purchased from a local market (Khuzestan, Iran). β‐carotene, Linoleic acid, ABTS (2,2′‐Azino‐bis (3‐ethylbenzothiazoline‐6‐sulfonic acid) diammonium salt), quercetin, gallic acid, and DPPH (2,2‐diphenyl‐1‐picrylhydrazyl) were purchased from Sigma‐Aldrich Co. Mannitol Salt Agar (MSA), Eosin Methylene Blue (EMB), Mueller Hinton Broth (MHB), Mueller Hinton Agar (MHA), Sabouraud Dextrose Broth (SDB), Sabouraud Dextrose Agar (SDA), and Plate Count Agar (PCA) were obtained from Merck Co.
7
Electrospun Antimicrobial PDLLA/PLGA Scaffolds
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The GMP-grade PDLLA (PURASORB® PDL 20, Mw = 230 kDa) and PLGA (LA50/GA50, PURASORB® PDLG 5010, Mw = 130 kDa) polymers were purchased from Corbion (Amsterdam, Netherlands). 2,2,2-Trifluoroetanol (TFE) organic solvent for electrospinning was purchased from Merck (Darmstadt, Germany). Phosphate-buffered saline (PBS, pH 7.4) was purchased from Fresenius Kabi (Bad Homburg vor der Höhe, Germany). Staphylococcus aureus (Gram-positive, ATCC 6538), Escherichia coli (Gram-negative, ATCC 8739), and normal human fetal osteoblasts (hFOB 1.19, ATCC CRL-11372TM) were purchased from American Type Culture Collection (ATCC). DMEM/Ham’s F12 medium, G418 disulfate salt, fetal bovine serum (FBS), penicillin–streptomycin solution, Live/dead Cell Double Staining Kit, paraformaldehyde, Triton-X100, Hoechst 33342, Mannitol Salt Agar, and MacConkey Agar were supplied by Sigma-Aldrich, St.Louis, MO, USA. AlexaFluorTM635 Phalloidin was purchased from ThermoFisher Scientific, Waltham, MA, USA, while fetal bovine serum (FBS) was purchased from PAN-Biotech (Aidenbach, Germany).
8
Bacterial Strain Characterization Protocol
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Clinical Escherichia coli and Staphylococcus aureus strains (Stellenbosch University Medical Microbiology, Tygerberg Hospital, Western Cape, South Africa) were inoculated in tryptic soy broth (3 g·L−1, 250 rpm, 37 °C, 24 h) within three days of isolation from a human host, and freeze cultures were prepared for storage after two passages (40% v·v−1 glycerol, −80 °C). All strains were inoculated directly from freeze culture stocks and passaged twice prior to experimental inoculation onto surfaces, after eight months of surface cleaning (3 g·L−1 tryptic soy broth, 250 rpm, 37 °C, 24 h). The clinical strains were not maintained as bench cultures at ambient temperatures, to prevent adaptation to laboratory conditions. Strains were characterized by plotting growth curves (3 g·L−1 tryptic soy broth, 37 °C, 250 rpm, 45 h). MacConkey agar (Sigma-Aldrich, Cape Town, South Africa) was used for the identification of E. coli. Mannitol salt agar (Sigma-Aldrich, Cape Town, South Africa) was used for the identification of S. aureus.
9
MRSA Cultivation and Biofilm Analysis
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The methicillin-resistant laboratory strain S. aureus ATCC 43300 was used in this study. Bacteria were stored in a cryovial bead preservation system (Roth, Karlsruhe, Germany) at −80 °C and cultivated on Brain-Heart Infusion (BHI; Sigma-Aldrich, Saint Louis, MO, USA) agar plates. Brain-Hearth Infusion broth was used for bacterial inoculum and phage production. Biofilm sonication fluids from larval infection experiments were plated on mannitol salt agar (Sigma-Aldrich, Saint Louis, MO, USA).
10
Isolation and Identification of Staphylococcus aureus from Milk
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A 10 µL of milk samples were plated on Mannitol Salt agar (Sigma-Aldrich, USA) and incubated at 37°C up to 48 hours. The appearance of the bacterial colonies was observed and recorded on 24 and 48 hours. Yellow bacterial colonies with yellow zones on the Mannitol Salt agar were picked and cultured on the Nutrient Agar supplemented with 7.5% of Sodium chloride. The suspected bacterial colonies were further examined using Gram staining and biochemical tests (haemolysis test, catalase test, oxidase test and coagulase test). Bacterial isolates that were gram positive, cocci shaped and showed biochemical characteristics identical to S. aureus were kept and cultured on the enrichment agar for further testing.
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