At d 56, 1 bird per replicate was selected and slaughtered for the collection of breast meat samples. The meat samples were stored in the refrigerator at 4 to 6oC for 3 days. After 3 days, the meat samples were removed and allowed to thaw and meat samples were analyzed using methods by Domanska and Rozanska (2003) . Using sterile spoons, 10g of breast meat was weighed aseptically into a sterile blender jar. About 90ml of sterile Butter eld's phosphate diluent or Buffered Peptone Water (BPW) was added and blended for two minutes. From the prepared homogenate about 10 serial dilutions were prepared by diluting 1 ml of homogenate with 9 ml of 0.1% peptone water and spread plated (0.1 ml) on a range of enumeration agar plates: Wilkins-Chalgren agar (Merck GmbH, Darmstadt, Germany) for estimation of clostridium counts, MacConkey agar (MAC) for coliforms; plate count agar (PCA) for heterotrophic bacteria (total aerobic plate counts); De Man Rogosa Sharpe agar (Merck KGaA, Foster City, California, United States) for estimation of lactobacilli count, and brilliant green agar (Merck Ltd, Mumbai, India) for estimation of salmonella count. The agar plates were incubated (Thermo Fisher Scienti c Inc., Waltham, MA, USA) at 370C for 24 hours. Microbial counts were expressed as colony-forming units (CFU) of microorganisms per gram of fresh sample.
Brilliant green agar
Manufactured by Merck Group
Sourced in India, Germany, United States
Brilliant green agar is a culture medium used for the selective isolation and enumeration of Salmonella species in food and clinical samples. It contains brilliant green dye, which inhibits the growth of Gram-positive bacteria and most Gram-negative bacteria, allowing Salmonella species to grow and be easily identified.
Automatically generated - may contain errors
Lab products found in correlation
De man rogosa sharpe agar, by Merck Group (1 mentions)
Potato dextrose agar (pda), by Thermo Fisher Scientific (1 mentions)
Macconkey agar, by Merck Group (1 mentions)
Ampicillin sodium salt, by Merck Group (1 mentions)
Eosin methylene blue agar, by Merck Group (1 mentions)
Mrs agar, by Merck Group (1 mentions)
5 protocols using brilliant green agar
1
Microbial Analysis of Chicken Breast Meat
2
Microbiological Viability Analysis of Chocolate
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Chocolate samples of each formulation were suspended in peptone water; serial dilutions were prepared and used for microbiological viability analysis. Enumeration of S. thermophilus CCM4757 in samples was determined by cultivating on M17 agar, after that incubated at 42 • C for 48 h, anaerobically with anaerobic kit. The counts were done on the 0 th , 7 th , 30 th , 60 th , 90 th , 120 th and 180 th days, were expressed as log CFU/g. Microbiological quality of chocolate samples was determined by counting yeasts, molds, and Salmonella. Brilliant Green Agar (Merck, Germany) was used Salmonella counting and incubated at 35 • C, for 24 h. The mold and yeast counting were performed in Potato Dextrose Agar (Oxoid, England) and incubated at 30 • C, for 120 h.
3
Phage MS2 Host Range and Plasmid Transfer
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E. coli J62 lac pro trp his pFlac::Tn3 (AmpR) and Salmonella Enteritidis P12510951 (link) from which the virulence plasmid had been eliminated52 (link) and the pFlac::Tn3 introduced (AmpR) were used as host for phage MS2. A rifampicin resistant mutant of the Salmonella strain (AmpR, Rif R) was used for the in vivo experiments. A collection of plasmid-containing wild-type AMR E. coli strains isolated from poultry37 (link) were selected for host range, phage MS2 sensitivity and plasmid loss experiments (Table S1 ). E. coli J62 Rif R was used as recipient strain for plasmid transfer rate assessment.
Luria Bertani (LB) broth and agar (1%, Sigma) was used for routine culture of the strains. Viable counts of E. coli J62 pFlac::Tn3 (AmpR) and AMR strains were determined by plating dilutions on MacConkey agar (Sigma) with or without antibiotics. The same was done for S. Enteritidis P125109 pFlac::Tn3 (AmpR, Rif R) counts, using Brilliant green agar (Sigma) supplemented with the appropriate antibiotic. For dilution purposes Phosphate-Buffered saline (PBS) buffer was used.
Luria Bertani (LB) broth and agar (1%, Sigma) was used for routine culture of the strains. Viable counts of E. coli J62 pFlac::Tn3 (AmpR) and AMR strains were determined by plating dilutions on MacConkey agar (Sigma) with or without antibiotics. The same was done for S. Enteritidis P125109 pFlac::Tn3 (AmpR, Rif R) counts, using Brilliant green agar (Sigma) supplemented with the appropriate antibiotic. For dilution purposes Phosphate-Buffered saline (PBS) buffer was used.
4
Quantification of S. enterica growth
5
Cecal Microbiome Enumeration in Rats
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Freshly collected cecal contents of rats were used immediately for enumeration of lactobacilli, bifidobacteria, E. coli, Salmonella and total aerobes using the conventional microbiological (spread plate) method. Briefly, 1 g of cecal content was suspended in 9 ml PBS and vortexed for 1 min. Samples were serially diluted in sterile diluents (0.5% peptone water in distilled water) and 100 μl of 10−4 to 10−6 dilutions were streaked on appropriate selective media for enumeration of different groups of bacteria. MRS agar was used for enumeration of lactobacilli, Bifidus Selective agar for bifidobacteria, Brain-Heart Infusion agar for total aerobes, Brilliant Green agar for Salmonella and Eosin Methylene Blue agar for E. coli (all media from Sigma, USA, except MRS from Merck, Germany). After incubation in appropriate conditions for each group of bacteria (72 h at 37°C in anaerobic condition for lactobacilli and bifidobacteria, and 48 h at 39°C in aerobic condition for Salmonella, E. coli and total aerobes), colonies on the plates were counted and microbial population was expressed as log CFU/g cecal content.
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