In addition to being plated on selective agar, 100 μL from each dilution of EC and PBS was also plated onto nutrient agar (DifcoTM, BD), MacConkey containing tetracycline (16 μg/mL, Sigma-Aldrich (SA), Saint Louis, MO, USA), ceftiofur hydrochloride (8 μg/mL, SA), or ampicillin (32 μg/mL, SA), or m-Enterococcus containing tetracycline (16 μg/mL), ceftiofur hydrochloride (8 μg/mL), or ampicillin (concentration of 16 μg/mL). Plates were incubated and counted; isolates were characterized and saved as described above. Isolates were considered resistant if they grew on their respective media containing antibiotics. This design is shown in Figure 1 . The antibiotic concentrations utilized in the plates are the CLSI breakpoints for the bacteria–antibiotic combination being tested. This was done to assess bacterial growth and resistance of human implication following veterinary administration of florfenicol. The MIC interpretive CLSI breakpoints can be found in Table 1 .
Macconkey
Manufactured by Merck Group
Sourced in Germany, United States
MacConkey is a selective and differential culture medium used for the isolation and identification of Gram-negative bacteria, particularly members of the Enterobacteriaceae family. It contains bile salts and crystal violet to inhibit the growth of Gram-positive bacteria, while allowing the growth of Gram-negative bacteria. The medium also contains lactose, and bacteria that can ferment lactose produce acid, which is detected by the pH indicator neutral red.
Automatically generated - may contain errors
Lab products found in correlation
Blood agar, by Merck Group (3 mentions)
Nutrient agar, by Merck Group (2 mentions)
Ceftiofur hydrochloride, by Merck Group (1 mentions)
Tetracycline, by Merck Group (1 mentions)
Ampicillin, by Merck Group (1 mentions)
Mrsaselect 2 agar, by Bio-Rad (1 mentions)
Cin agar, by Merck Group (1 mentions)
Gram staining, by Merck Group (1 mentions)
Api kits, by bioMérieux (1 mentions)
50 ml falcon tube, by Sarstedt (1 mentions)
Fastdna spin kit for soil, by MP Biomedicals (1 mentions)
Cefotaxime, by Merck Group (1 mentions)
Brain heart infusion broth, by Thermo Fisher Scientific (1 mentions)
Vitek 2 gn id card, by bioMérieux (1 mentions)
Ciprofloxacin, by Merck Group (1 mentions)
Tryptone bile x glucuronide agar, by Merck Group (1 mentions)
Vitek 2 compact, by bioMérieux (1 mentions)
Simmons citrate agar, by Merck Group (1 mentions)
Baird parker, by Merck Group (1 mentions)
Api staph kit, by bioMérieux (1 mentions)
18 protocols using macconkey
1
Assessing Antibiotic Resistance in Bacteria
2
Endometritis Bacterial Profiling in Cows
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Forty uterine secretion samples were collected from Holstein and Jersey cows from clinical cases of endometritis. The samples were collected in sterile containers. Prior to bacteriological examination, the samples were mixed with sterile distilled water in a vortex shaker for 10–15 min, and divided into two halves. One half was used for bacterial counting, and the other half was used for identification of organisms after direct inoculation on different agar plates, including BHI, nutrient, blood, and MacConkey (Sigma-Aldrich, St. Loius, MO, USA) agar plates.
3
Bacterial Two-Hybrid Assay for UvrB-UvrC Interaction
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Interaction between full-length UvrB and UvrC proteins was probed using the Bacterial Adenylate Cyclase Two Hybrid (BACTH) system in E. coli (48 (link)). For this purpose, the uvrB and uvrC genes were cloned into the pEB354 and pEB355 to be expressed as fusion proteins respectively with the T25 and T18 fragments of adenylate cyclase. The reporter strain BTH101 was co-transformed with the two plasmids, one encoding the T18-bait fusion protein and the other encoding the T25-prey fusion protein (or empty plasmids expressing T18 or T25 alone as controls), and transformants were selected on LB agar plates supplemented with ampicillin and kanamycin incubated at 30°C for 48 h. Five 3 ml LB cultures containing ampicillin, kanamycin and 0.5 mM IPTG, were then inoculated with individual colonies from each transformation plate and incubated in a shaking incubator overnight at 30°C. The next day, 3 μl from each culture were spotted in duplicate on lactose-containing MacConkey (Sigma) agar plates supplemented with ampicillin and kanamycin, which were placed at 30°C for 48 h. Lactose fermenting bacteria, resulting from bait-prey interaction, are red in color. In contrast, the absence of interaction results in beige colonies.
4
Fecal Microbiome Quantification in Lambs
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On the last day of the experiment (09:00 h) fecal samples were collected (10 g/lamb) in sterile collection tubes by rectal stimulation and using latex gloves. The tubes were maintained at 4°C during transport to the laboratory. One gram of each sample was diluted in a Labcon tube with 9 mL peptonized solution (8.5%). It was then homogenized in vortex for serial dilutions of 10−1 to 10−12. Each dilution was seeded in triplicate 100 μL by the striate method in Petri dishes containing selective medium for coliforms (Mac Conkey, Sigma-Aldrich, Munich, Germany) or for lactic acid bacteria (de Man, Rogosa and Sharpe, Sigma-Aldrich, Germany) and incubated 24 and 48 h at 37°C, respectively. After incubation, colony-forming units were counted to estimate the microbial population, and the data were expressed with the base 10 logarithm function (log10). The material and the culture media used in this phase were sterilized 15 min in autoclave (Lab-Med, LMV40, Mexico City, Mexico) at 121°C and 15 psi, and the dilutions and bacterial seeding were performed in a laminar flow hood (Labconco, Logic A2 800, Kansas City, MO, USA).
5
Murine Wound Infection Model
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Bacteria was grown as described above, normalized to 106 CFU/10 μL, and used to infect wounds of C57BL/6 mice (Male, 7–8 weeks old; InVivos, Singapore) as previously described [92 (link)]. Briefly, the animals were anesthetized with 3% isoflurane. Dorsal hair was shaven and fine hair was removed after the application of Nair cream (Church and Dwight Co, Charles Ewing Boulevard, USA) and shaved using a scalpel blade. The skin was disinfected with 70% ethanol and a full-thickness wound was created with a 6 mm biopsy punch (Integra Miltex, New York, USA). The wounds were inoculated with 10 μL of the respective inoculum (E. coli, 1–2 x 106 CFU; S. aureus 1–2 x 106 CFU; Mixed 1–2 x 106 CFU each). Thereafter, the wound site was sealed with a transparent dressing (Tegaderm 3M, St Paul Minnesota, USA). At the indicated timepoints, mice were euthanized, and the wounds were excised and homogenized in 1 mL PBS. Viable bacteria in the wound homogenates were enumerated by plating onto selective media for E. coli (MacConkey; Merck Singapore) and S. aureus (MRSASelect II Agar; Biorad USA).
6
Microbial Isolation and Identification
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The samples were collected in two sterile microtubes for culture and PCR. For the PCR test 1000 µL of PBS (phosphate buffer salts) was added in to the microtubes containing the patient’s direct samples and the tubes were stored at -70°C. Microbiological culture of the samples was performed in the bacteriology lab using a suitable culture media such as MacConkey (Merck, Germany) agar and selective agar for isolation of organism and biochemical differential test reactions for identification of the isolates.
7
Characterization of Shigella Strains
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S. sonnei ATCC® 9290 and S. flexneri ATCC 12022 standard strains were obtained from Iranian National Center for Genetic and Biologic Resources. Eighteen clinical isolates were also collected for assessment of host-specific antibacterial assay. Additionally, Staphylococcus aureus (S. aureus) ATCC 43300 was employed as the control. The strains were subcultured onto the blood agar (Merck, Germany) and MacConkey (Merck, Germany, for Shigella species) media and kept at -20°C until further experiments. Furthermore, 18 clinical isolates of both species were collected and identified using biochemical tests.
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S. sonnei ATCC® 9290 and S. flexneri ATCC 12022 standard strains were obtained from Iranian National Center for Genetic and Biologic Resources. Eighteen clinical isolates were also collected for assessment of host-specific antibacterial assay. Additionally, Staphylococcus aureus (S. aureus) ATCC 43300 was employed as the control. The strains were subcultured onto the blood agar (Merck, Germany) and MacConkey (Merck, Germany, for Shigella species) media and kept at -20°C until further experiments. Furthermore, 18 clinical isolates of both species were collected and identified using biochemical tests.
8
Bacterial Pathogen Screening in Diarrhea
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Bacteriological differential media were used to screen for diarrheagenic bacteria including Salmonella spp., Shigella spp., Yersinia spp., and E. coli. To screen and further identify E. coli, stool samples were initially plated on MacConkey (Merck®, Darmstadt, Germany) agar. Five individual lactose fermenter isolates that had metallic shine on Eosin Methylene Blue (EMB) agar (Merck, Darmstadt, Germany) were subsequently processed with biochemical assays for E. coli confirmation. Salmosyst Broth (Merck, Darmstadt, Germany) with supplements (Merck, Darmstadt, Germany) was used for isolation of Shigella and Salmonella species and CESY Broth (Merck, Darmstadt, Germany) was used for recovery of Yersinia at 37°C for 24 hours, respectively (Figure 1 ). Bacteria able to grow in selective media were subsequently cultured in either Salmonella-Shigella (SS) agar (Scharlau®, Sentmenat, Spain) or CIN-Agar (Merck, Darmstadt, Germany) for further selection of Salmonella, Shigella, or Yersinia species.
9
Isolation of Enteric Pathogens from Livestock
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Samples were aseptically collected from calves, foals, and piglets by rectal swabbing, placed in sterile tubes, and transferred to the laboratory in a cool box within 3 h of collection. After arrival in the Microbiology Laboratory in the University of Agricultural Sciences and Veterinary Medicine Cluj, rectal swabs were transferred into separate tubes containing 2 mL nutrient broth and cultured at 37°C for one d following the steps stated in the ISO 16654:2001 protocol (26 ). Briefly, each sample was inoculated on MacConkey (Merck, Darmstadt, Germany) agar plates and incubated at 37°C overnight.
10
Isolation and Identification of Pseudomonas Isolates
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Swab samples were inoculated on blood, MacConkey (Merck, Germany) and nutrient agar (Merck, Germany) and incubated at 37°C for 24 hours. Colonies that produced pyoverdin, pyocyanin and pyorubin pigments were transferred to nutrient agar and subcultured more than one time to obtain pure cultures. The isolates were identified using conventional biochemical tests such as motility, oxidase, catalase, citrate utilization, gelatinase liquefaction, urease production, nitrate reduction, alkaline protease production, triple sugar iron agar, oxidative-fermentative, indole, lecithinase production and hemolysin production.
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