After processing for 4 min in washing water with an FC concentration of 10 ppm, the sample was dewatered using an alcohol-sterilized salad spinner and sealed in a polyethylene terephthalate box using polyvinyl chloride cling film [29] . The samples were stored at 4 °C and analyzed on days 0, 3, and 7. Pathogens were analyzed as described in Section 2.3.2 . For naturally present microbes, 0.1 mL suspension was surface plated on Rose Bengal agar (Hopebio) and incubated at 30 °C for 3 d to quantify molds and yeasts (M&Y); 1 mL bacterial suspension was pour-plated onto plate count agar (Hopebio) and incubated at 37 °C for 2 d to obtain the aerobic mesophilic counts (AMCs) and at 7 °C for 10 d to obtain the aerobic psychrophilic counts (APCs).
Plate count agar
Manufactured by Hopebio
Sourced in China
Plate count agar is a solid culture medium used for the enumeration of viable microorganisms in a sample. It provides a nutritious environment for the growth and development of bacterial colonies, enabling the quantification of microbial populations.
Automatically generated - may contain errors
Lab products found in correlation
Rose bengal agar, by Hopebio (8 mentions)
Xylose lysine deoxycholate agar, by Hopebio (4 mentions)
Modified sorbitol macconkey agar, by Hopebio (4 mentions)
Violet red bile agar, by Hopebio (3 mentions)
Du730, by Beckman Coulter (1 mentions)
Dna extraction kit, by BioTeke (1 mentions)
Potato dextrose agar (pda), by Hopebio (1 mentions)
Stomacher 400, by Seward Medical (1 mentions)
Taq pcr mastermix, by Tiangen Biotech (1 mentions)
Urea, by Merck Group (1 mentions)
Proteinase k, by Merck Group (1 mentions)
Mrs agar, by Hopebio (1 mentions)
Boric acid, by Sangon (1 mentions)
Sodium chloride (nacl), by Sangon (1 mentions)
2 thiobarbituric acid, by Sangon (1 mentions)
18 protocols using plate count agar
1
Pathogen and Microbial Analysis of Washed Samples
2
Microbiological Analysis of Food Samples
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Microbiological analysis was performed on days 0, 3, and 7. A sample (15 g) was placed in a stomacher bag containing 135 mL of 0.85 % NaCl and homogenized for 2 min, followed by serial dilution. Then, 0.1 mL of the diluted bacterial suspension was surface-plated on modified sorbitol MacConkey agar (Hopebio, Qingdao, China) or xylose lysine deoxycholate agar (Hopebio) and incubated at 37 °C for 24 h to count E. coli O157:H7 and Salmonella Typhimurium, respectively. For naturally present microbes, 0.1 mL of suspension was surface-plated on Rose Bengal agar (Hopebio) and incubated at 30 °C for 3 days to quantify molds and yeasts (M&Y). In addition, 1 mL of the suspension was pour-plated onto plate count agar (Hopebio) and incubated at 37 °C for 2 days to obtain the aerobic mesophilic count (AMC). All results are expressed as log CFU/g.
3
Microbial Analysis of Disinfected E. coli O157:H7
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E. coli O157:H7, AMC, aerobic psychrotrophic count (APC), coliforms, and molds and yeasts (M&Y) were analyzed immediately after acid disinfection and at the end of the storage period (day 5). At the sampling points, 10 g was sampled and each piece was divided into four parts to ensure full contact with the NaCl solution. Next, 5 g of the sample was placed in an Erlenmeyer flask containing 70 mL of 8.5 g/L NaCl solution and shaken for 3 min at 260 rpm to prepare a 15-fold dilution, after which 6 mL of the suspension was added to 34 mL of 8.5 g/L NaCl solution to prepare a 100-fold dilution [2 (link)]. Other dilutions were prepared as needed. A 1 mL dilution was pour-plated onto (1) plate count agar (Hopebio) and incubated at 37 °C for 2 days for the measurement of AMC, as well as at 7 °C for 10 days for the measurement of APC and (2) violet red bile glucose agar (VRBGA; Hopebio) and incubated at 37 °C for 1 day for the measurement of coliforms. Moreover, a 0.1 mL dilution was surface-plated on (3) rose Bengal agar (Hopebio) and incubated at 30 °C for 3 days for the measurement of M&Y; (4) SMAC agar (Hopebio) and incubated at 37 °C for 1 day for the measurement of E. coli O157:H7; and (5) Listeria chromogenic agar (Land Bridge) and incubated at 37 °C for 1 day for the measurement of L. monocytogenes. The results are expressed as microbial reductions (log CFU g−1).
4
Bacterial Counts in Mutton Sausage
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The bacterial counts of mutton sausage samples were determined according to the method of Chen et al. (2019) (link). Total aerobic counts were detected using Plate Count Agar (PCA; Qing Dao Hope Bio-Technology Limited Corporation, Qing Dao City, China) after incubation at 37°C for 48 h. Presumptive lactic acid bacteria counts were determined on de Man Rogosa Sharpe Agar (MRSA, Qing Dao Hope Bio-Technology Limited Corporation, Qing Dao City, China) after incubation at 30°C for 48 h. Staphylococci counts were cultured on Mannitol Salt Agar (MSA, Qing Dao Hope Bio-Technology Limited Corporation, Qing Dao City, China) after incubation at 32°C for 17 h.
5
Microbial Enumeration in Food Samples
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After disinfection, the samples (15 g) were transferred to a sterilized stomacher bag, diluted 1:15 in 225 mL of sterilized 0.85% NaCl solution and homogenized in a stomacher for 90 s. For E. coli and L. monocytogenes, 0.1 mL of diluted bacterial suspension was surface-plated on SMAC agar and Listeria chromogenic agar, respectively, and microbial counts were obtained after a 24-h incubation period at 37 °C. For naturally present microbial taxa, 0.1 mL of the diluted bacterial suspension was surface-plated on rose Bengal agar (Hopebio) and incubated at 30 °C for three days to quantify molds and yeasts (M&Y). Additionally, 1 mL of the dilution was pour-plated onto plate count agar (Hopebio) and incubated at 37 °C for 2 days to obtain aerobic mesophilic counts (AMC) and at 7 °C for 10 days to obtain aerobic psychrotrophic counts (APC).
6
Microbial Counts in Salmon Filets
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The total mesophilic bacterial counts (MBC), total psychrotrophic bacteria count (PBC), and H2S-producing bacterial counts (HSPBC) of salmon filets were determined according to Yu et al. (28 (link)). In brief, 25 g meat sample were taken from each group under sterile condition and homogenized with 225 mL of 0.85% sterilized saline water with a stomacher blender for 1 min. From this dilution, a series of dilutions (1:10) were prepared in 0.1% peptone, and 0.1 mL of each dilution was dispensed onto the appropriate medium. Plate count agar (Qingdao Hope Bio-Technology Co., Ltd., Shandong, China) was used for determining total mesophilic bacteria count (MBC) (48 h incubation at 30 °C) and total psychrotrophic bacteria count (PBC) (10 d incubation at 4 °C). The H2S-producing bacteria count was determined by counting black colonies on Iron agar (Qingdao Hope Bio-Technology Co., Ltd., Shandong, China) after incubation at 25°C for 72 h. Each dilution was performed in three parallels.
7
Multiplex EuNP-based LFIA–RPA for Foodborne Pathogens
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The strains used in this study are listed in Table 1 . Listeria monocytogenes (ATCC19111), Vibrio parahaemolyticus (ATCC33847), and Escherichia coli O157:H7 (ATCC35150) were chosen as the reference strains for all inoculation experiments of multiplex EuNP-based LFIA–RPA. Listeria monocytogenes strains were grown on buffered Listeria enrichment broth (BLEB) (Thermo Fisher Scientific Inc., Waltham, MA, USA). Vibrio parahaemolyticus strains were grown on alkaline peptone water (APW, Hopebio, Qingdao, China) supplemented with 3% NaCl. Escherichia coli O157:H7 strains were grown on lauryl tryptose (LST) broth (Thermo Fisher Scientific Inc., Waltham, MA, USA). All strains were incubated overnight at 35 °C for 20–24 h. The bacterial culture was used for the extraction of the genome. According to the Bacteriological Analytical Manual (BAM), the cell number was calculated using plate count agar (Hopebio, Qingdao, China).
The total DNA was extracted using a DNA extraction kit (Bioteke Corporation, Beijing, China) in accordance with the manufacturer’s instructions. The concentration and quality of the DNA were measured using a spectrophotometer (DU730, Beckman Coulter, Burea, CA, USA). The genomic DNA was stored at −20 °C until use.
The total DNA was extracted using a DNA extraction kit (Bioteke Corporation, Beijing, China) in accordance with the manufacturer’s instructions. The concentration and quality of the DNA were measured using a spectrophotometer (DU730, Beckman Coulter, Burea, CA, USA). The genomic DNA was stored at −20 °C until use.
8
Microbial Analysis of Food Samples
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Samples were analyzed at 0, 3, and 7 days. A 25-g sample was homogenized with 225 mL sterile NaCl solution for 1.5 min in a stomacher bag. Then, the suspension was serially diluted. The suspension (0.1 mL) was surface-plated on modified sorbitol MacConkey agar (Hopebio), Listeria chromogenic agar (Land Bridge, Beijing, China), and xylose lysine deoxycholate agar (Hopebio) to analyze E. coli O157:H7, L. monocytogenes, and Salmonella Typhimurium, respectively, and incubated for 24 h at 37 °C. For naturally present microbes, 0.1 mL of the diluted bacterial suspension was surface-plated on Rose Bengal agar (Hopebio) and incubated at 30 °C for 3 days to quantify molds and yeasts (M&Y). In addition, 1 mL of the suspension was pour-plated onto plate count agar (Hopebio) and incubated at 7 °C for 10 days to obtain the aerobic psychrotrophic count (APC) and or at 37 °C for 2 days to obtain the aerobic mesophilic count (AMC). All results are expressed as log CFU/g.
9
Rapid Enumeration of E. coli and Salmonella
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Eight samples were randomly selected from the package, transferred to a sterile stomacher bag, diluted 1:9 (w/v) in sterile 0.85% NaCl solution, and homogenized in a stomacher for 2 min. Then, 1 mL of the diluted bacterial suspension was spread-plated on modified sorbitol MacConkey agar (Hopebio) and xylose lysine deoxycholate agar (Hopebio) and incubated for 24 h at 37 °C to analyze E. coli O157:H7 and S. Typhimurium, respectively. For naturally-present microbes, 1 mL of the bacterial suspension was pour-plated in plate count agar (Hopebio) and incubated at 37 °C for 2 d to obtain the AMC, and 1 mL was pour-plated in rose bengal agar (Hopebio) and incubated at 28 °C for 5 days to quantify the M&Y.
10
Microbial Quantification Protocol
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The sample and the sterilized 0.85% NaCl were added into a stomacher bag at a ratio of 1:9 (w/v) and homogenized for 2 min to obtain a bacterial suspension. The diluted bacterial suspension (1 mL) was pour-plated into plate count agar (Hopebio, Qingdao, China) and incubated for 2 days at 37 °C to analyze the aerobic mesophilic count (AMC). For molds and yeasts (M&Y), 1 mL diluted suspension was pour-plated into rose bengal agar (Hopebio) and incubated for 5 days at 28 °C. The results were expressed as log CFU/g.
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