For phospholipase detection, the method proposed by Samaranayake et al13 (link) was used with modification. Aliquots of the yeast suspension (10 μl) were added to wells punched onto the surface of the egg yolk agar medium (HiMedia). The diameter of the precipitation zone around the well was measured after incubation at 37°C for two days. Phospholipase activity (Pz value) was determined as previously described13 (link). When Pz=1, no phospholipase activity was detected in the strain. C. albicans ATCC 10231 was used as a positive control.
Egg yolk agar medium
Manufactured by HiMedia
Sourced in India
Egg yolk agar medium is a culture medium used for the isolation and identification of Clostridium perfringens, a spore-forming anaerobic bacterium. It contains egg yolk, which allows the detection of lecithinase production, a key characteristic of C. perfringens.
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Lab products found in correlation
Blood agar medium, by HiMedia (1 mentions)
Triple sugar iron agar, by HiMedia (1 mentions)
Phosphate buffer saline (pbs), by HiMedia (1 mentions)
Starch agar medium, by HiMedia (1 mentions)
Sim medium, by HiMedia (1 mentions)
Lauria bertani broth, by HiMedia (1 mentions)
Coomassie brilliant blue, by HiMedia (1 mentions)
Nutrient agar, by HiMedia (1 mentions)
Sodium acetate, by HiMedia (1 mentions)
5 protocols using egg yolk agar medium
1
Phospholipase Activity Assay in Yeast
2
Isolation and Characterization of Staphylococcus aureus
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Bacterial cultures were collected from the region of the middle nasal meatus and from the throat of the 70 inpatients by use of sterile cotton-tipped wire swabs. The specimens were transported immediately to the laboratory in transport media and then inoculated on blood agar. Bacterial cultures were identified morphologically and biochemically by standard laboratory procedures according to the Manual of Methods for General Bacteriology in the bacteriological laboratory of Sumy State University. We isolated 50 Staphylococcus aureus strains. Each culture underwent Gram staining and was tested for production of catalase, free coagulase, yellow pigment, mannitol fermentation, growth on high salt concentration, and lipase production on egg yolk agar medium (Hi Media, Mumbai).
3
Staphylococcus aureus Identification Assays
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The bacterial strains that were confirmed as Staphylococcus aureus by species specific 16S rRNA screening were further analyzed by several microbiological diagnostics tests including mannitol fermentation and growth on high salt concentration, gelatin hydrolysis, urea hydrolysis, protease activity on milk agar medium, lipase production on egg yolk agar medium (HiMedia, Mumbai), and hydrolysis of esculin by standard methods [23 (link)–26 (link)]. Hemolytic activity was determined on sheep blood agar (15 mL of 5% sheep blood in Trypticase soy agar overlaid on 10 mL of blood agar base) according to Rodgers et al. [27 (link)].
4
Biochemical Characterization of S. aureus
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The bacterial strains which were confirmed as S. aureus by species-specific 16S rRNA were further evaluated by different biochemical tests including mannitol fermentation and growth on high salt concentration, gelatin hydrolysis, urea hydrolysis, protease activity on milk agar medium, lipase production on egg yolk agar medium (HiMedia, Mumbai), and hydrolysis of esculin by standard method [16 (link)–19 (link)] and congo red agar (CRA) test [20 (link)].
5
Soil Microbial Diversity in Nepal
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This study was carried out in Central Campus of Technology Hattisar Dharan and Regional Agricultural Research Station (RARS) Tarhara from October 2018 to March 2019. Soil samples were collected from different altitudes ranging from Itahari (374 ft.), Tarhara (418 ft.), Dharan (1272 ft.) and Vedetar (5140 ft.) of Koshi Zone, Nepal. From each geographical region 25 different soil samples were collected and in total 100 different soil samples from four regions were collected. Soil samples weighing 10gm was collected in sterile plastic bags from 3-5cm depth and transported into the laboratory at 4°C. Materials used in this experiment were Sodium acetate (HiMedia, India), Lauria Bertani (LB) broth (HiMedia, India). Nutrient Agar (HiMedia, India), Coomassie brilliant blue (HiMedia, India), Phosphate buffer saline (HiMedia, India), Grams staining reagents (HiMedia, India), MR-VP medium and reagents (HiMedia, India), Indole medium (HiMedia, India), Citrate agar medium (HiMedia, India), Gelatin agar medium (HiMedia, India), Starch agar medium (HiMedia, India), Egg yolk agar medium (HiMedia, India), Blood agar medium (HiMedia, India), Carbohydrate fermentation broth (HiMedia, India), SIM medium (HiMedia, India), Triple sugar iron agar (HiMedia, India).
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