Several dilutions of the fish samples were prepared. At each sampling interval, 10 g of fish fillets was mixed thoroughly with a sterile saline solution (0.85% NaCl) for 1 min to prepare the initial dilution (10−1). Serial dilutions were plated on plate count agar (Merck, Germany) for total aerobic bacterial counts (TABC), followed by incubation at 35 °C for 48 h.40 Samples for total anaerobic bacterial counts (TANBC) were plated on plate count agar and anaerobically incubated 48 h at 35 °C.40 Enterobacteriaceae counts were carried out using violet red bile glucose agar (oxide), with the plates incubated for 24 h at 37 °C.41 Staphylococci counts were performed by spreading 0.1 ml of a suitable dilution on the surface of a Baird Parker agar plates complemented with egg yolk and potassium tellurite solution and incubating for 48 h at 35 °C.41 Counts of mold and yeast were performed by spreading 0.1 ml of a suitable dilution on the surface of rose bengal chloramphenicol agar plates and aerobically incubated at 25 °C for four days.42 Finally, psychotropic bacteria were detected using plate count agar and incubated at 5 °C for seven days.41 All microbiological results were expressed as log10 CFU g−1.
Plate count agar
Manufactured by Merck Group
Sourced in Germany, United States, Italy, China
Plate count agar is a microbiology culture medium used for the enumeration of viable microorganisms in a sample. It supports the growth of a wide range of bacteria and provides a reliable method for determining total bacterial counts.
Automatically generated - may contain errors
Lab products found in correlation
Potato dextrose agar (pda), by Merck Group (10 mentions)
Violet red bile agar, by Merck Group (6 mentions)
Mrs agar, by Merck Group (5 mentions)
Violet red bile glucose agar, by Merck Group (4 mentions)
Sabouraud dextrose agar, by Merck Group (4 mentions)
Tryptic soy broth (tsb), by Merck Group (4 mentions)
Hydrochloric acid (hcl), by Merck Group (4 mentions)
Sodium chloride (nacl), by Merck Group (4 mentions)
Peptone water, by Merck Group (4 mentions)
Man rogosa sharpe agar, by Merck Group (3 mentions)
Macconkey agar, by Merck Group (3 mentions)
Malt extract agar, by Merck Group (3 mentions)
Mannitol salt agar, by Merck Group (3 mentions)
Linoleic acid, by Merck Group (3 mentions)
Nutrient agar, by Merck Group (3 mentions)
Mueller hinton agar (mha), by Merck Group (3 mentions)
Sodium hydroxide (naoh), by Merck Group (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Methanol, by Merck Group (3 mentions)
Cetrimide fucidin cephaloridine agar, by Merck Group (2 mentions)
104 protocols using plate count agar
1
Microbiological Analysis of Fish Samples
2
Fecal Microbiome Analysis in Rats
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On days 0, 7, 14 and 21, a pooled fecal sample from the rats in each subgroup was taken. Just after collection, each subgroup’s fecal sample was weighed and homogenized in sterile phosphate buffer saline (PBS), and then, tenfold serial dilution was made in PBS. Aerobic and anaerobic microorganisms were cultured in Plate Count Agar (Merck, Germany). Lactic acid bacteria (LAB) were cultured in MRS (de Man, Rogosa, Sharpe) agar (Merck, Germany). Coliforms were cultured in VRBL (Violet Red Bile Lactose) agar (Merck, Germany). LAB and anaerobic bacteria were incubated at anaerobic condition. All plates were incubated at 37°C for 48 h. The microflora enumeration was expressed as CFU/gr of feces. In order to count bacterial spores in fecal samples, the homogenized samples in PBS were put into a water bath at 80°C for 10 min, and then cooled immediately to help the spore germination. Then the samples were serially diluted in PBS and cultured in Plate Count Agar (Merck, Germany). The plates were incubated at 37°C for 48 h. The spore enumeration was expressed as spores/gr. All the samples were cultured in duplicate (17 (link), 19 ).
3
Gut Microbiome Modulation in Salmonella Infection
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One day before and days 1, 3, 5 and 7 after intragastric gavage of S. Typhimurium, a pooled fecal sample from the rats in each subgroup was taken. Just after collection, each sample was weighed, homogenized in sterile phosphate buffer saline (PBS) (pH: 7.2), and tenfold serial dilutions were made in PBS. Aerobic and anaerobic microorganisms were cultured in Plate Count agar (Merck, Germany). LAB were cultured in MRS (de Man, Rogosa, Sharpe) agar (Merck, Germany) and the Coliforms were cultured in VRBL (Violet Red Bile Lactose) agar (Merck, Germany). Anaerobes and LAB medium plates were placed in an anaerobic jar. All plates were incubated at 37 °C for 48 h. The micro ora enumeration was expressed as CFU/gr . In order to count bacterial spores in fecal samples, the homogenized samples in PBS were put into a water bath at 80 °C for 10 min and cooled immediately. Then the samples were serially diluted in PBS and cultured in Plate Count agar (Merck, Germany). The plates were incubated at 37 °C for 48 h. The spore enumeration was expressed as spores/gr. All the samples were cultured in duplicate [23 -25] .
4
Microbiological Analysis of Beef Burgers
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Samples (30 g) were aseptically taken from each beef burger and homogenized with peptone water (0.1%) in a Lab-Blender for 3 min to have a final dilution of 1:10. Serial decimal dilutions were made using the same diluent and then plated in duplicate for bacterial counts. Aerobic mesophilic bacteria were determined on plate count agar (Merck, Darmstadt, Germany) after 48 h incubation at 30 °C. Mold and yeast were counted on acidified potato dextrose agar (Merck, Darmstadt, Germany) after 48 h incubation period at 28 °C. Coliform group was determined on MacConkey agar (Merck, Darmstadt, Germany) after 48 h incubation at 37 °C. Psychrotrophs were determined on plate count agar (Merck, Darmstadt, Germany) after ten-day incubation at 7 °C [19] .
5
Enumeration of Microbial Loads in Fruit Salad
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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)].
6
Evaluating Wound Contraction and Bacterial Load
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The wound contraction rate was assessed with the help of a transparent paper over the wound site and tracing it52 ,53 (link). The total bacterial count was also measured with the help of plate count agar (Merck KGaA, Darmstadt, Germany). In summary, 0.1 g of the wound sample was crushed, minced, and diluted and cultured on plate count agar. The cultured plates were incubated at 37 °C for 48 h and the total bacterial count was calculated.
7
Microbiological Analysis of Fish Fillets
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Ten grams of the fish fillets were mixed with 90 ml of sterile saline solution (0.09% w/v) and homogenized in a stomacher for 2 min. Then, the serial dilution was made for the following microorganism determinations: (a) Viable mesophilic bacteria were determined using plate count agar (Merck) after incubation for 24 hr at 37°C, (b) psychotropic bacteria were determined using Man, rogosa and sharpe (Macromedia PTY‐LTD) after 10 d of incubation at 4–7°C, (c) lactic acid bacteria were determined by MRS agar (Merck) for 48 hr at 37°C, and (d) Pseudomonas were determined using cetrimide agar (CA) for 2 days of incubation at 25°C.
8
Antimicrobial and Antioxidant Properties Evaluation
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Saline peptone water, Brain Heart Infusion (BHI), Tryptic Soy Broth (TSB) supplemented with Yeast, Plate Count Agar (PCA), Tryptone Sulphite Neomycin (TSN), Dextrose Sabouraud (DS), and Mannitol Egg Yolk Polymyxin (MYP) mediums were obtained from Merck KGaA, (Darmstadt, Germany). Acetonitrile; amphotericin B solution (250 μg/mL); 2,2′-Azinobis 3-ethylbenzothiazoline-6-sulphonic acid (ABTS); Fetal Bovine Serum (FBS); glacial acetic acid; hydrochloric acid; 6-hydroxy 2,5,7,8-tetramethyl-2-carboxylic acid (Trolox); L-glutamine solution (200 mM); MEM (Minimum Essential Medium); methanol; penicillin (10,000 U/mL); pure phenolic compounds (caffeic acid, catechin, epicatechin, epicatechin gallate, ethyl gallate, gallic acid, gentisic acid, p-coumaric acid, procyanidin B1, procyanidin B2, protocatechuic acid, syringic acid, t-resveratrol, t-piceid, and vallinic acid); streptomycin solution (P/S; 100 mg/mL); and 2,4,6-Tris (2-pyridyl)-S-triazine (TPTZ) were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Folin–Ciocalteu (FC) reagent, 70% (v/v), iron (III)-chloride acid (FeCl3), iron (II)-sulphate (FeSO4), sodium acetate (NaC2H3O2), and sodium carbonate (Na2CO3) were purchased from Panreac Quimica S.L.U. (Barcelona, Spain). Malvidin-3-glucoside was obtained from Estrasynthese (Genay, France).
9
Synthesis and Characterization of TiO2 Nanoparticles
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TiO2 nanoparticles (anatase) with purity of more than 99% and particle size of 10–25 nm were obtained from US Research Nanomaterials, Inc. Na‐alginate was obtained from Behin Azma Co. Nutrient broth, Violet Red Bile Agar (VRBA), Cetrimide fucidin cephaloridine agar (CFC agar), De Man, Rogosa, Sharpe agar (MRS), Listeria Chrom agar, and plate count agar (PCA) were purchased from Merck Chemical Co.
10
Antimicrobial Activity of Spice Extracts
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The freshly caught shrimp were obtained from a local market in Fasa (Fars province, Iran) and brought to the laboratory with ice. After properly washing shrimps, they were kept at −18°C until testing. All culture media, including plate count agar (PCA), cetrimide fucidin cephaloridine agar (CFC), de Man‐Rogosa‐Sharpe agar (MRS), and violet red bile glucose (VRBG), as well as peptone water (PW), Tween 20 and Tween 80 were provided by Merck Company (Merck, Darmstadt, Germany). Sodium alginate and CaCl2 were also obtained from Sigma Company (Sigma‐Aldrich, Germany). Zataria multiflora and C. cyminum EOs were purchased from Zardband Pharmaceuticals and Tabibdaru companies (Iran), respectively. Distilled water was used in all tests.
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