The TBC and total numbers of L. monocytogenes and S. Typhimurium in the CBF were analyzed on the 1st, 3rd, 5th, 7th, 10th, and 12th days of storage. For each sampling day, the 25 g chicken samples were homogenized in 225 mL 0.1% buffered peptone water using a stomacher (Seward, 400 Circulator, Worthing, UK) for 1 min. Homogenized samples were then serially diluted using 0.1% buffered peptone water. They were spread-plated on plate count agar (for TBC, Merck Millipore 105463, Burlington, MA, USA), Wilson-Blair bismuth sulfite agar (for S. Typhimurium count, Merck Millipore 100191, Burlington, MA, USA), and Oxford base agar with the Listeria Selective Supplement (for L. monocytogenes count, Merck Millipore 107004 with 107006, Burlington, MA, USA). All plates were incubated at 37 °C for 48 h [33 ,34 ,35 ]. The numbers of presumptive colonies on each sampling day were recorded as log colony-forming units/g sample (log cfu/g).
400 circulator
Manufactured by Seward Medical
Sourced in United Kingdom
The 400 Circulator is a laboratory equipment designed to provide precise and consistent temperature control for various applications. It features a digital display that allows for easy monitoring and adjustment of the temperature. The 400 Circulator is capable of maintaining temperatures within a specified range to support various experimental and research processes.
Automatically generated - may contain errors
Lab products found in correlation
Buffered peptone water (bpw), by Liofilchem (2 mentions)
Rappaport vassiliadis broth, by Thermo Fisher Scientific (2 mentions)
Api 20e strip, by bioMérieux (2 mentions)
Xylose lysine deoxycholate (xld), by Thermo Fisher Scientific (2 mentions)
Anaerocult c, by Merck Group (1 mentions)
Mrs agar, by Merck Group (1 mentions)
Salmonella antiserum, by Denka Seiken (1 mentions)
Cm0509, by Thermo Fisher Scientific (1 mentions)
Cm0469, by Thermo Fisher Scientific (1 mentions)
Dneasy powerfood microbial kit, by Qiagen (1 mentions)
Nucleospin food kit, by Macherey-Nagel (1 mentions)
Api 20e diagnostic strips, by bioMérieux (1 mentions)
Sorbitol macconkey agar, by Liofilchem (1 mentions)
12 protocols using 400 circulator
1
Microbiological Enumeration of Chicken
2
Meat Sample Homogenization and Dilution
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Twenty‐five grams of each meat sample (for chicken a mixture of meat from neck, breast, wings, and legs) was added into 225 ml of Buffered Peptone Water (Liofilchem, Teramo, Italy) and homogenized using a stomacher (400 Circulator; Seward, London, UK). Dilutions of 10 to 10 were realized from the stock solution according to ISO methods (ISO6887‐2 2004 ).
3
Isolation and Characterization of Lactic Acid Bacteria
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Amount of 25 g of each sample was aseptically weighted and homogenized with 225 ml of sodium citrate using a stomacher (Seward Stomacher 400 Circulator, UK). Samples were subsequently diluted (1:10) using sterile peptone water and 0.1 ml from each dilution was sub-cultured on Man Rogosa and Sharpe (MRS) agar (Merck, Germany) used for isolating lactic acid bacteria [6 ]. Plates were incubated at 30°C and 37°C for mesophilic lactic acid bacteria and at 42°C for thermophilic lactic acid bacteria for 48 h. MRS agar plates were incubated anaerobically using the gas pack systems (Anaerocult C, Merck, Germany). To differentiate homo- and heterofermentative strains, all the isolates were tested for gram reaction, catalase production, and carbohydrates fermentation (homofermentatives showed ribose +, sorbitol +, raffinose ±, lactose +, galactose +, maltose +, melibiose ±, rhamnose −, xylose ±, manose +, salicine +, sorbose +, glucose +, and sucrose + pattern, heterofermentatives showed ribose +, sorbitol ±, raffinose ±, lactose ±, galactose ±, maltose +, melibiose ±, rhamnose ±, xylose −, manose +, salicine ±, sorbose ±, glucose +, and sucrose±pattern). Cimon citrate test was also conducted as a differential test [7 ,8 ].
4
Isolation and Identification of E. coli in Fish Samples
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25 g of raw or cooked fish samples was homogenized for 2 min. in a sterile bag containing 225 ml of buffered peptone water (0.1%) (Lab M, UK) using a stomacher (Seward Stomacher 400 circulator, UK). All samples were inoculated onto eosin methylene blue medium (EMB) agar and incubated at 37°C for 24 h. Suspected colonies on EMB agar showed a green metallic sheen appearance and were subcultured on MacConkey agar medium and nutrient agar to get a pure colony and were incubated at 37°C for 24 h (Gupta et al., 2013 ). Colonies of E. coli on EMB suggest a green metallic sheen appearance. Colonies suspected to be E. coli were subjected to biochemical identification (Alexander et al., 2010 (link)).
The suspected result from the above media was inoculated into nutrient agar and tested by different biochemical tests: Indole test, Methyl red test, Simon citrate test, Triple sugar iron agar (TSI) test, and Urease test.
The suspected result from the above media was inoculated into nutrient agar and tested by different biochemical tests: Indole test, Methyl red test, Simon citrate test, Triple sugar iron agar (TSI) test, and Urease test.
5
Antimicrobial Effects of BITC and γ-CD-BITC on Cooked Chicken
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Fresh chicken breast was cut into cube blocks (approximately 25 g). The sterile cooked chicken breast samples were then obtained by sterilizing the cube blocks for 20 min at 121 °C. Samples of the BITC (or γ-CD-BITC) at 0.25 mmol/L and the bacterial suspension (about 107 CFU/mL) were added, followed by homogenization for 120 s in a stomacher (400 Circulator, Seward, England). The inoculated samples were stored by using aseptic bags at 10, 15, 20, and 25 °C, respectively. At each sampling time point, three chicken samples were tested for microbiological examination.
The data of S. aureus in the cooked chicken breast treated with BITC and γ-CD-BITC (basing on BITC concentration) were modeled using the modified Gompertz model [6 (link),32 (link)] as followed:
where y0, ymax, and Y(t) are the bacterial population counts in a natural logarithm of S. aureus at initial, maximum, and time (Log CFU/g); μmax is the maximum specific growth rate (Log CFU/g/h); and λ is the lag time duration (h).
The data of S. aureus in the cooked chicken breast treated with BITC and γ-CD-BITC (basing on BITC concentration) were modeled using the modified Gompertz model [6 (link),32 (link)] as followed:
where y0, ymax, and Y(t) are the bacterial population counts in a natural logarithm of S. aureus at initial, maximum, and time (Log CFU/g); μmax is the maximum specific growth rate (Log CFU/g/h); and λ is the lag time duration (h).
6
Salmonella Detection Protocol
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Each sample was placed in a stomacher bag containing 90 ml of buffered peptone water (CM0509, Oxoid) and homogenized for 1 minute (400 Circulator, Seward, London, UK) before being incubated at 37°C for 24 hours, in accordance with ISO 6579-1 (2017) . Transferring 0.1 ml of nonselective pre-enrichment to 10 ml of Rappaport-Vassiliadis broth (CM0866, Oxoid) and incubating at 42°C for 24 hours. A loopful of the selective enrichment broth was streaked on xylose lysine deoxycholate agar (XLD) (CM0469, Oxoid) agar and incubated for 24 hours at 37°C. Salmonella colonies that exhibited a red coloration with black centers on XLD agar were identified both morphologically and biochemically, as described by Quinn et al. (2002 ). Serologically, Salmonella isolates were serologically identified using the Kauffman-White-Le Minor system (Grimont and Weill, 2007 ) to detect Somatic (O) and Flagellar (H) antigens with Salmonella antiserum (DENKA SEIKEN Co., Japan).
7
Comparative Analysis of Food DNA Extraction
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A sample of 20 g was suspended in 180 ml of sterile Buffered Peptone Water (BPW) and homogenized using a Stomacher 400 Circulator (Seward, United Kingdom) for 5 min at 300 rpm. Then, 1 ml of the solution was transferred to 2-ml tubes and centrifuged for 1 min at 14,000 rpm. The supernatant was discarded, and the pellet was resuspended in each examined kit lysis buffer. Three commercial food DNA extraction kits were used: (a) blackPREP Food DNA I Kit (Analytik Jena AG) (BP), (b) DNeasy® PowerFood® Microbial Kit (MoBio Laboratories Inc., Carlsbad, CA, United States) (MB) και, and (c) Nucleospin® Food Kit (MACHEREY-NAGEL) (NS), for bacterial and yeast/fungi metagenomic DNA extraction. Additionally, three different cell lysis principles were applied before the DNA extraction process (Table 2 ):
Then the DNA extraction process was performed according to each kit manufacturer’s instructions. The extracted DNA was stored at −20°C until processing.
Then the DNA extraction process was performed according to each kit manufacturer’s instructions. The extracted DNA was stored at −20°C until processing.
8
Vibrio Detection in Shrimp Samples
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The shrimps (5 g) were homogenized with alkaline peptone water (45 ml) (lab M, UK) containing 1% NaCl for 2 min by a stomacher (Seward 400 circulator, UK). Likewise, water (1 ml) was vortexed in alkaline peptone water (9 ml), and then incubated at 35 ± 2°C for 24 h. Enrichment cultures were platted onto thiosulphate-citrate-bile salts-sucrose (TCBS- lab M, UK), where yellow and green colonies were counted as Vibrio-like colonies. Selected colonies were streaked onto TSA slants supplemented with 1% NaCl. After incubation at 35 ± 2°C/24 h, the isolates were tested biochemically with oxidase test and API 20E diagnostic strips (BioMérieux, France) [31 (link), 32 ].
9
Enumeration of Bacterial Colonies in Rocket Leaves
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10 g of rocket leaves was added to the 90 ml preparation of MRD and placed in a stomacher (400 Circulator; Seward, Worthing, UK) and shaken for 120 s (230 rpm) to create a 10−1 dilution (w/v). 1 ml of the homogenised inoculum was sampled and serially diluted into the 9 ml MRD preparation to obtain 10−2, 10−3, 10−4, 10−5, 10−6, and 10−7. 1 ml of each respective solution was added to 15 ml nutrient agar (45–50 °C) in petri dishes, using the pour plate technique. Plates were swirled to mix evenly. Inoculated plates were allowed to cool at room temperature (∼22 °C) until the liquid solidified, and incubated at 30 °C in inverted condition. After 72 ± 3 h, the number of colonies per plate were counted using a colony counter. Bacterial numbers for each sample were estimated in colony forming units (cfu·g−1).
10
Isolation and Identification of E. coli
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The isolation of Escherichia coli strains was carried out according to the guidelines of ISO 4832: 1951 (F). Briefly, 25 g of each sample were added into 225 mL of buffered peptone water (BPW) (Liofilchem, Teramo, Italy) and homogenized using a stomacher (400 Circulator, Seward, London, UK). This step is followed by cascade dilution, which could often go up to 10−7 dilution depending on the type of sample. A loopful of these diluents was streaked onto sorbitol MacConkey agar (Liofilchem, Teramo, Italy) and incubated at 44 °C overnight. After incubation, suspected colonies were confirmed by biochemical tests including indole, methyl red, Voges–Proskauer and citrate [31 ]. The confirmed colonies were conserved in tubes with 1 mL brain heart infusion broth containing 15% (v/v) glycerol for further analysis at 4 °C.
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