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Bacteriological agar

Manufactured by Acumedia
Sourced in Brazil

Bacteriological agar is a solidifying agent derived from red seaweed. It is used in the preparation of culture media for the growth and isolation of microorganisms in a laboratory setting.

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2 protocols using bacteriological agar

1

Evaluating Phosphate Solubilization Capacity

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Modified Pikovskaya agar medium was used to determine the phosphate solubilisation capacity (Nopparat et al. 2009 ). The base solution (BS) was composed of (NH4)2SO4 (0.5 g), KCl (0.2 g), MgSO4.H2O (0.1 g), MnSO4.H2O (0.004 g), FeSO4.7H2O (0.002 g), NaCl (0.2 g), D-glucose (10 g, Sigma-Aldrich), yeast extract (0.5 g, Kasvi, Brazil), bacteriological agar (18 g, Acumedia, USA), and distilled water (900 mL). The pH was adjusted to 6.8. The first phosphate solution (PS1) was composed of xanthan gum (0.5 g, Sigma-Aldrich), Ca3(PO4)2 (0.5 g, β-tricalcium phosphate, Sigma-Aldrich) and distilled water. For the second solution (PS2), calcium phosphate was replaced with bone meal (0.5 g). All solutions were autoclaved (15 min at 121°C), mixed (BS+ PS1 and BS + PS2; pH adjusted to 6.6), and transferred to petri dishes.
To evaluate the growth of endophytes under different phosphorous sources, 6-mm plugs of each fungus were inoculated in petri dishes with BS+PS1 and BS+PS2 and incubated at 28°C for 7 days. The diameter of the halo surrounding the fungal colony and the colony diameter were measured daily using a graduated ruler (cm). The relative solubilisation efficiency was calculated as the diameter of solubilisation halo/diameter of colony × 100, using the mean of five replicates.
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2

Mouse Stomach Dissection and Analysis

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The mice were fasted for 2 h prior to sacrifice by cervical dislocation, and blood samples were taken by heart puncture and kept on ice. Serum samples were separated as soon as the samples arrived at the lab by centrifugation for 10 minutes at 11,500 × g at room temperature and were stored at – 80°C until use. The mouse autopsy started from the first cut to remove the ventral skin. A second cut across the midline opened the abdominal cavity, and two subsequent cuts removed the sternum. Terminal blood was collected by cardiac puncture. The stomach was extracted from the abdomen by cutting off 2–3 mm from the esophagus to the gastroduodenal junction. Before opening the stomach, the pH values were measured. The stomach was then opened along the long curvature, emptied of its contents, and weighed. The stomach was divided into two parts, including the forestomach, corpus, and antrum. One part was fixed in 4% PFA (HistoLab, Sweden) for histological examination of the tissue. The mucous layer of second part of the stomach was scraped off with a sterile blade and placed into transport medium (2 g casamino acid (Difco), 2 g peptone (VWR), 0.4 g yeast extract (Merck), 0.32 g bacteriological agar (Acumedia), 0.04 g Lcysteine (Merck), 0.2 g glucose, 28 ml glycerol, 1 g sodium chloride (Merck), and 240 mL Milli-Q (Millipore) filtered water (pH 7.0) for quantitative culture.
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