Kefir samples (10 mL) were dispersed with 90 mL of sterile sodium thiosulfate solution (0.2% wt/vol) and homogenized for 1 min using a stomacher (Smasher, AES Chemunex, Bruz, France). Further dilutions were made using Ringer solution (Merck, Darmstadt, Germany). Lactobacilli counts were determined on de Man Rogosa Sharpe agar (Merck) after incubation at 30°C under anaerobic conditions for 5 d (García Fontán et al., 2006) . Lactococci counts were enumerated on M17 Agar (Merck) at 30°C for 3 d (Irigoyen et al., 2005) . Leuconostoc spp. counts were determined on de Man Rogosa Sharpe agar (Merck) incorporated with vancomycin hydrochloride (Sigma-Aldrich, St Louis, MO) at 30°C for 3 d (García Fontán et al., 2006) and yeasts were grown on yeast extract glucose chloramphenicol agar (Merck) at 25°C for 3 d (Magra et al., 2012) .
De man rogosa sharpe agar
Manufactured by Merck Group
Sourced in Germany, United States
De Man Rogosa Sharpe Agar is a culture medium used for the selective isolation and enumeration of lactobacilli. It provides the nutrients and conditions required for the growth of these bacteria.
Automatically generated - may contain errors
Lab products found in correlation
Anaerocult a, by Merck Group (2 mentions)
Vancomycin hydrochloride, by Merck Group (2 mentions)
M17 agar, by Merck Group (2 mentions)
Eosin methylene blue agar, by Merck Group (1 mentions)
Violet red bile dextrose agar, by Merck Group (1 mentions)
Mannitol salt phenol red agar, by Merck Group (1 mentions)
Rose bengal chloramphenicol agar, by Thermo Fisher Scientific (1 mentions)
Blood agar, by Thermo Fisher Scientific (1 mentions)
Macconkey agar, by Merck Group (1 mentions)
Potato dextrose agar (pda), by Merck Group (1 mentions)
Mrs sorbitol agar, by Merck Group (1 mentions)
M17 medium, by Merck Group (1 mentions)
Brilliant green agar, by Merck Group (1 mentions)
Yeast extract glucose chloramphenicol agar, by Merck Group (1 mentions)
Ringer solution, by Merck Group (1 mentions)
11 protocols using de man rogosa sharpe agar
1
Enumeration of Kefir Microbiota
2
Enumeration of Broiler Chicken Gut Microbiota
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At 35 d of age, 6 broiler chickens per treatment were chosen and sacrificed. The intestinal segment was removed immediately and 3 g of fresh digesta from ileum was collected sterilely. The samples were put on ice until they were transported to the laboratory for enumeration of microbial populations. Each sample was serially diluted from 1/10 to 1/107 in sterilized physiological saline solution (NaCl 85%). Then 0.1 mL of each diluted sample was plated onto the following media. E. coli was cultured on eosin methylene blue agar (Merck, Darmstadt, Germany) at 37 °C for 24 h. Lactobacilli bacteria were counted on de Man, Rogosa, sharpe agar (Merck, Darmstadt, Germany) after incubation for 48 to 72 h at 37 °C.
3
Breast-fed Infant Gut Microbiome
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Feces samples (n = 38) of breast-fed infants in Songklanakarind Hospital were collected with the necessary approval from the Ethics Committees of the Faculty of Medicine, Prince of Songkla University (REC.61-064-4-2). Infants were enrolled according to the following criteria: age < 6 months, exclusively received breast milk with predominant LAB strains, vaginal delivery, healthy infants, mothers without present or past underlying adverse medical conditions, and full-term pregnancy. The feces were immediately cultured on de Man Rogosa Sharpe agar (Merck Millipore, Darmstadt, Germany) at 37°C for 48 h under anaerobic conditions. After incubation, each of the isolated colonies were picked and stored at −80°C in BHI broth with 30% glycerol until testing.
4
Screening Antifungal Activity of Lactic Acid Bacteria
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Qualitative analysis of the antifungal activity was carried out in accordance with Strom’s method [16 ]. The medium used for the growth of A. flavus was PDA with a mold spore concentration of 106 spores/mL. Lactic acid bacteria isolates were chosen to detect the presence of antifungal activity with a streak plate method on de man rogosa sharpe agar (Merck). Aspergillus flavus was inoculated on the medium using the spread plate method, and the LAB isolates were subsequently inoculated by the streak plate method. The results of inoculation were grown under anaerobic conditions for 24 h at a temperature of 37°C. After incubation, if a clear zone was formed in the medium, the LAB isolates were considered to have antifungal activity and were reselected for the next screening.
5
Enumeration of Lactobacillus in Fermented Plant-Based Milk
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Microbiological analysis included the determination of the cell number of Lactobacillus strains in the bean-based plant substitute of fermented milk by the pour plate method with two parallel independent replicates of each of the analyzed beverage. The De Man–Rogosa–Sharpe agar (Merck KGaA) was used for the analysis. The Petri dishes with inoculations were incubated at 37 °C for 72 h under anaerobic conditions using an anaerobic jar equipped with Anaerocult A (Merck KGaA). After the completion of incubation, the grown colonies were counted. For colony counting, the Petri dishes in which the number of grown colonies was in the range of 30–300 colonies were chosen. The final results were converted to the number of colony-forming units per gram of the product (CFU/g), and then converted to logarithm (log10 CFU/g).
6
Enumeration of Microbial Populations in Food
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Samples of 25 g were added to 225 mL physiological saline (0.85 NaCl%) and homogenized in a stomacher (Lab Stomacher, London, UK) for 1 min. In determining the number of lactic acid bacteria, de Man Rogosa Sharpe Agar (Merck, Darmstadt, Germany) was used and incubation was carried out at 30 °C for 48 h under anaerobic conditions (Anaerocult A, Merck, Darmstadt, Germany). Mannitol Salt Phenol Red Agar (Merck) was used for the enumeration of Micrococcus/Staphylococcus, and the plates were incubated aerobically at 30 °C for 48 h. Enterobacteriaceae were determined on Violet Red Bile Dextrose Agar (Merck), and the plates were incubated at 30 °C for 48 h under anaerobic conditions (Anaerocult A, Merck) [23 ].
7
Tarhana Fermentation Microbial Enumeration
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Tarhana samples were taken every 24 h during fermentation. The samples were transferred aseptically into 0.1% peptone water and the dilutions were mixed or spread on different media and incubated under the appropriate conditions. For mesophilic and thermophilic lactic acid bacteria, de Man-Rogosa-Sharpe agar (Merck, KGaA, Darmstadt, Germany) containing 0.02% sodium azide was incubated for 48 h at 30°C and 42°C, respectively and for yeasts and moulds, Rose Bengal Chloramphenicol Agar (Oxoid, England) containing supplements (Oxoid, England) was incubated at 30°C for 48 h [19 (link)].
8
Microbiological Analysis of Piglet Fecal and Digesta Samples
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Microbiological analyses were performed on faecal samples (collected daily from one piglet per pen) and digesta samples collected from piglets that were killed at the end of the experiment. Approximately 1 g of faeces and 10 g of digesta were suspended in a pre-reduced salt medium(18 ) and homogenised in a laboratory paddle blender (Seward Stomachers® 80 Biomaster; Lab System) for 2 min. Serial 10-fold dilutions were then prepared in the same medium and samples (0·1 ml) were plated on selective media. Haemolytic E. coli and coliform bacteria were enumerated on blood agar (Oxoid) and MacConkey agar (Merck), respectively, after aerobic incubation at 37°C for 1 d, whereas lactic acid bacteria (LAB) were enumerated on de Man–Rogosa–Sharpe agar (Merck) after anaerobic incubation at 37°C for 3 d. The number of E. coli and LAB is presented as the LAB:haemolytic E. coli ratio or the LAB:total coliform bacteria ratio.
9
Microbial Enumeration in Kefir Samples
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Ten-gram kefir samples were aseptically taken. Sterilized Ringer's solution at a dilution of 1:9 (w/v) was added, and the samples were homogenized for 3 min in a stomacher Lab-Blender 400 (London, UK). The serial decimal dilutions were prepared in buffered peptone water and plated for bacterial counts.
Lactobacillus was counted on de Man Rogosa and Sharpe medium (Merck 1.10660) at 30 °C under anaerobic conditions (5% CD 2 ) for 3 d. Lactococcus was counted on M17 medium from Merck at 37 °C under anaerobic conditions for 2 d. Leuconostoc Spp. was counted on de Man Rogosa Sharpe Agar (Merck) incorporated with vancomycin hydrochloride (Sigma-Aldrich, St Louis, MD, USA) at 30 °C for 3 d. Lactobacillus acidophilus was counted on MRS-sorbitol Agar (Merck 1.10660) at 37 °C under anaerobic conditions for 24-48 h. (Tomar et al., 2018) . Yeasts were cultured on potato dextrose agar (Merck, 1.10130) (pH 3.5) with 10% added tartaric acid (Akarca et al., 2016) (link).
Lactobacillus was counted on de Man Rogosa and Sharpe medium (Merck 1.10660) at 30 °C under anaerobic conditions (5% CD 2 ) for 3 d. Lactococcus was counted on M17 medium from Merck at 37 °C under anaerobic conditions for 2 d. Leuconostoc Spp. was counted on de Man Rogosa Sharpe Agar (Merck) incorporated with vancomycin hydrochloride (Sigma-Aldrich, St Louis, MD, USA) at 30 °C for 3 d. Lactobacillus acidophilus was counted on MRS-sorbitol Agar (Merck 1.10660) at 37 °C under anaerobic conditions for 24-48 h. (Tomar et al., 2018) . Yeasts were cultured on potato dextrose agar (Merck, 1.10130) (pH 3.5) with 10% added tartaric acid (Akarca et al., 2016) (link).
10
Microbial Analysis of Chicken Breast Meat
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At d 56, 1 bird per replicate was selected and slaughtered for the collection of breast meat samples. The meat samples were stored in the refrigerator at 4 to 6oC for 3 days. After 3 days, the meat samples were removed and allowed to thaw and meat samples were analyzed using methods by Domanska and Rozanska (2003) . Using sterile spoons, 10g of breast meat was weighed aseptically into a sterile blender jar. About 90ml of sterile Butter eld's phosphate diluent or Buffered Peptone Water (BPW) was added and blended for two minutes. From the prepared homogenate about 10 serial dilutions were prepared by diluting 1 ml of homogenate with 9 ml of 0.1% peptone water and spread plated (0.1 ml) on a range of enumeration agar plates: Wilkins-Chalgren agar (Merck GmbH, Darmstadt, Germany) for estimation of clostridium counts, MacConkey agar (MAC) for coliforms; plate count agar (PCA) for heterotrophic bacteria (total aerobic plate counts); De Man Rogosa Sharpe agar (Merck KGaA, Foster City, California, United States) for estimation of lactobacilli count, and brilliant green agar (Merck Ltd, Mumbai, India) for estimation of salmonella count. The agar plates were incubated (Thermo Fisher Scienti c Inc., Waltham, MA, USA) at 370C for 24 hours. Microbial counts were expressed as colony-forming units (CFU) of microorganisms per gram of fresh sample.
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