Under aseptic conditions, 10 g of fruit salad was homogenized with 90 mL of sterile 0.85% physiological saline solution to establish decimal dilutions using the same diluent. After initial experimentation, three suitable serial dilutions were chosen, and the spread plate technique was employed to inoculate the respective culture media, with incubation periods per specific requirements. Total mesophilic bacteria (TMAB) was enumerated at 28 °C for 48 h using Plate Count Agar (PCA, Rahway, NJ, USA, Merck). Total psychrotrophic bacteria (TPB) enumeration took place at 4 °C for 10 days using PCA. Total yeast and mold (TYM) was generated at 28 °C for 3–5 days using Yeast Glucose Agar (YGC, Merck). Finally, total Enterobacteriaceae (TEN) enumeration was performed at 37 °C for 24 h using Violet Red Bile Glucose Agar (VRBG, Merck). The microbial load was expressed as log colony-forming units (CFU) per gram of fruit salad [15 (link)].
Violet red bile glucose agar vrbg
Manufactured by Merck Group
Sourced in Germany
Violet Red Bile Glucose Agar (VRBG) is a culture medium used for the selective isolation and enumeration of Enterobacteriaceae in food and water samples. It contains crystal violet and bile salts to inhibit the growth of Gram-positive bacteria, while glucose serves as the fermentable carbohydrate. The agar allows for the differentiation of Enterobacteriaceae based on colony morphology and color.
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Lab products found in correlation
Yeast glucose agar ygc, by Merck Group (2 mentions)
Plate count agar pca, by Merck Group (2 mentions)
Plate count agar, by Merck Group (1 mentions)
M17 agar, by Merck Group (1 mentions)
Anaerocult c, by Merck Group (1 mentions)
Mrs agar, by Merck Group (1 mentions)
Sodium thiosulfate, by Merck Group (1 mentions)
Acetic acid, by RCI Labscan (1 mentions)
Potassium iodide, by Merck Group (1 mentions)
1 1 3 3 tetramethoxypropane, by Merck Group (1 mentions)
2 propanal, by Merck Group (1 mentions)
Palmitic acid, by Merck Group (1 mentions)
Potassium dihydrogen phosphate, by Merck Group (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
Trichloroacetic acid, by Merck Group (1 mentions)
Thiobarbituric acid, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Merck Group (1 mentions)
Chloroform, by Merck Group (1 mentions)
Phenolphthalein, by Merck Group (1 mentions)
5 protocols using violet red bile glucose agar vrbg
1
Enumeration of Microbial Loads in Fruit Salad
2
Microbial Analysis of Fruit Salad
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For microbial analysis, 10 g of the fruit salad (2.5 g from each fruit) was homogenized (MIX 2, AES Laboratoire, France) with 90 mL of a saline solution (0.85% w/v sodium chloride (NaCl)). The mixture was then appropriately diluted to achieve the desired microbial count. From these dilutions, samples were plated on specific agar media: Plate Count Agar (PCA, Merck) for total mesophilic aerobic bacteria, Yeast Glucose Agar (YGC, Merck) for total yeast and mold, and Violet Red Bile Glucose Agar (VRBG, Merck) for total Enterobacteriaceae count. After incubation at 30°C for 48 h, 25°C for 3–5 days, and 37°C for 24 h, respectively, the results were reported as log colony‐forming unit (CFU)/g, following the methodology outlined by Lacivita et al. (2021 (link)).
3
Enumeration of Yoghurt Microbiota
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Bacteria were enumerated in yoghurt samples by the plate count method. Yoghurt samples in 1 mL aliquots were combined with 9 mL of sterile saline solution (NaCl 0.9%, w/v). Coliforms were enumerated at 30 °C on Violet Red Bile Glucose agar (VRBG) (Merck, Warsaw, Poland). Starter cultures were enumerated on selective media based according to IDF standards [31 ]. Lactobacillus delbrueckii ssp. bulgaricus strains were isolated on MRS agar (Merck, Warsaw, Poland) and adjusted to pH 5.4 by microaerophilic incubation in an anaerobic jar with Anaerocult C (Merck, Warsaw, Poland) at 37 °C for 72 h. Streptococcus thermophilus strains were isolated on M17 agar (Merck, Warsaw, Poland), adjusted to pH 7.2, by aerobic incubation at 37 °C for 48 h. Microbial counts were expressed as log10 of colony-forming units (cfus) per mL of yoghurt.
4
Microbial Enumeration in Food Samples
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Ten grams of each representative sample were taken and placed in a sterile plastic bag with 90 mL of peptone water (0.1%) with 0.85% NaCl. After 2 min in a stomacher blender (Stomacher Colworth 400, Seward, UK), appropriate decimal dilutions were pour-plated (1 mL) on the following media: Plate Count Agar (PCA) (Merck, Darmstadt, Germany) for the total viable count (TVC) (30 °C for 72 h); De Man, Rogosa, Sharpe Agar (MRS) (Merck, Darmstadt, Germany) for lactic acid bacteria (LAB) (30 °C for 3–5 days); and Violet Red Bile Glucose Agar (VRBG) (Merck, Darmstadt, Germany) for Enterobacteriaceae (37 °C for 24 h). All microbial counts were converted to logarithms of colony-forming units per gram (Log cfu/g), following the methodology used by Triki et al. [20 (link)].
5
Chitin-Chitosan Powder Characterization
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The food grade CS powder with 200 mesh particle size had molecular weight of 8.97 × 105 DA and 80% degree of deacetylation. It was purchased from Sinudom Agriculture Products Co., Ltd. (Surat Thani, Thailand). NI was provided by Shandong Freda Biotechnology Co., Ltd. (Shandong, China). CO (Syzygium aromaticum, Lin), with eugenol content in 70–80% range (manufacturer's data) was purchased from Thai China Flavors and Fragrances Industry Co., Ltd. (Bangkok, Thailand). The chemical agents used were analytical grade: chloroform, dipotassium hydrogen phosphate, phenolphthalein, methanol, potassium iodide, trichloroacetic acid, hydrochloric acid, 2-propanal, sodium hydroxide, sodium thiosulfate, thiobarbituric acid, potassium dihydrogen phosphate, anhydrous sodium sulfate, (Merck, Darmstadt, Germany), acetic acid (Lab-Scan, Bangkok, Thailand), glycerol (Vidhyasom, Thailand), Tween 80 (Labchem, Australia), palmitic acid, and 1,1,3,3-tetramethoxypropane (Sigma-Aldrich, St. Louis., MO, USA). The media for microbiological analyses were analytical grade: plate count agar; peptone; deMan, Rogosa, and Sharpe (MRS) agar; and violet red bile glucose agar (VRBG) (Merck, Darmstadt, Germany).
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