The following O157 strains were used in peptide susceptibility assays: (i) EDL933 (ATCC 43895; stx1+, stx2+, eae+, hlyA+; American Type Culture Collection/ATCC, Manassas, VA) and (ii) 86–24 (NADC 6103; stx2+, eae+). O157 strains EDL933 and 86–24 displayed similar results in initial peptide susceptibility assays, and due to EDL933 having both stx genes, only strain EDL933 was used in subsequent assays. Bacterial freezer stocks were streaked individually on Luria-Bertani (LB) Lennox agar (Sigma-Aldrich, St. Louis, MO) and incubated overnight at 37˚C. Bacterial cultures were prepared by growing each isolate from a single colony in 5 mL LB broth (Sigma-Aldrich) per assay for 20 h at 37˚C with constant shaking at 150 rpm. Assay isolates were streaked on Difco sorbitol MacConkey agar (BD Biosciences, Franklin Lakes, NJ) containing 4-methylumbelliferyl-β-d-glucuronide (100 mg/L; Sigma-Aldrich) (SMAC-MUG) or SMAC-MUG supplemented with cefixime (50 μg/L), potassium tellurite (2.5 mg/L), and vancomycin (40 mg/L) (SMAC-CTMV) agar [32 (link), 33 (link)]. Plates were read after overnight incubation at 37˚C and colonies that did not ferment sorbitol or utilize 4-methylumbelliferyl-β-d-glucuronide (non-fluorescent under UV light) were confirmed to be O157 via latex agglutination (E. coli O157 latex, Oxoid Diagnostic Reagents, Oxoid Ltd., Hampshire, UK) [32 (link), 33 (link)].
Edl933
Manufactured by American Type Culture Collection
Sourced in United States
The EDL933 is a laboratory equipment product used for the cultivation and storage of bacterial strains. It provides a controlled environment for the growth and maintenance of microbial cultures, including those associated with the E. coli O157:H7 serotype.
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Lab products found in correlation
5 protocols using edl933
1
Characterization of O157 Escherichia coli Strains
2
Bacterial Antimicrobial Susceptibility Assay
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The bacterial strains used were enterohemorrhagic Escherichia coli (E. coli) O157:H7 [ATCC 43895, EDL933], Pseudomonas aeruginosa (P. aeruginosa PAO1), methicillin-sensitive Staphylococcus aureus (MSSA 6538) [ATCC 6538], and methicillin-resistant Staphylococcus aureus (MRSA) strain [ATCC BAA-1707]. Bacterial experiments were performed at 37°C in Luria–Bertani (LB) medium for E. coli, P. aeruginosa, and S. aureus MSSA strains and in LB medium containing 0.2% glucose for S. aureus MRSA strain. For phenotypic assays, stationary phase cells were reinoculated into LB medium at an initial turbidity of 0.05 (OD600). All reagents and materials used in this study were purchased from Sigma-Aldrich (St Louis, MO, USA) and used as received.
3
Antimicrobial Whey Protein Film
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Whey protein concentrate (WPC) 80% (http://pdfood.com.br/Whey %20Protein%20-%201.pdf; Arla Food Ingredients S.A, Martinez, Argentina) and glycerol (Gly) (Cicarelli, San Lorenzo, Argentina) were used to prepare film forming solutions. Five soy sauces (SS) were acquired from different local manufacturers (SS1, SS2, SS3, SS4 and SS5). Mueller-Hinton broth and agar were purchased from Britania (Buenos Aires, Argentina). All other reagents were of analytical grade. The microorganisms used were Escherichia coli O157:H7 strain EDL933 (ATCC 43895) obtained from Área Bacteriología Clínica (Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Argentina), Salmonella Typhimurium (ATCC 14028) provided by Dr. Susana Checa (Instituto de Biología Molecular y Celular de Rosario, IBR-CONICET), and Listeria monocytogenes (ATCC 19115) kindly donated by Dr. Ana Bessone (Laboratorio Americano, Rosario, Argentina).
4
Induction and Propagation of E. coli Phages
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E. coli strains BAA2326 (containing phage BAA2326), EDL 933 (containing phage 933W), K802, and MG1655 were obtained from the American Type Culture Collection (Manassas, VA, USA). Bacterial cultures were grown with agitation in Luria Broth (LB) at 37 °C. Phages were obtained by induction of the respective host strains with mitomycin C (1 µg/mL). λ and 434 were obtained from our collection. BAA2326 and 933W lysogens were constructed as described previously [29 ].
All plasmids were propagated in E. coli strain K802. BAA2326 repressor was purified from the E. coli strain BL21(DE3) pLysS (Novagen, Madison, WI, USA) bearing a plasmid that directs its overexpression. Primers for site-directed mutagenesis and making other templates for DNA binding experiments were obtained from Integrated DNA Technologies (Coralville, IA, USA).
All plasmids were propagated in E. coli strain K802. BAA2326 repressor was purified from the E. coli strain BL21(DE3) pLysS (Novagen, Madison, WI, USA) bearing a plasmid that directs its overexpression. Primers for site-directed mutagenesis and making other templates for DNA binding experiments were obtained from Integrated DNA Technologies (Coralville, IA, USA).
5
Bacterial Culture Protocols for E. coli
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Bacterial cultures were grown with agitation either in Luria Broth (LB), or M9 + 0.08% glucose with or without added monovalent salts. Media was supplemented with 50 μg/mL chloramphenicol where appropriate. Escherichia coli strains EDL933, BAA2326, and MG1655 were obtained from the American Type Culture Collection (Manassas, VA, USA) The MG1655recA strain, which bears the recA938::cat allele, was created by P1 transduction [29 (link)]. The Shiga toxin-encoding phages 933W and BAA2326 were obtained by induction of the respective host strains. λ was obtained from a constructed W3110:λ lysogen previously reported [3 (link)], and the 434 was from our collection [29 (link)].
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