Infected animals (10 animals per group) were monitored every 6 to 10 h for clinical signs until the animal was lethargic or until day 7 postinfection. An animal still alive at 7 days postinfection was considered a survivor, and this time point was also a surrogate for the endpoint of the experiment. The bacterial loads in blood were measured at 24 h postinfection and at the time when the animal was lethargic or at the end of the experiment for survivors. At 24 h postinfection, ∼25 to 50 μl of blood was collected from the tail vein and at the time of animal expiration by cardiac puncture under deep anesthesia (5.0% [vol/vol] fluothane [Zeneca Pharmaceuticals, United Kingdom] over oxygen at a rate of 1.5 to 2 liters/min). The animal was euthanized by cervical dislocation, and the lungs were removed, weighed, placed into 10 ml of PBS, and immediately homogenized. Pneumococcal CFU were tested in the lung homogenates on blood agar base plates (Oxoid) in serial decimal dilutions and then converted to CFU per milligram of lung tissue.
Blood agar base plates
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
Blood agar base plates are a type of growth medium used in microbiology laboratories. They provide a nutrient-rich environment that supports the growth of a wide range of microorganisms, particularly those that require blood or blood components for their growth. The agar base is supplemented with defibrinated blood, typically sheep or horse blood, to create a solid culture medium.
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Lab products found in correlation
2 protocols using blood agar base plates
1
Pneumococcal Infection Monitoring in Animals
2
Isolation and Identification of Vancomycin-Resistant Enterococci from Meat Samples
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One-hundred grams of each sample were aseptically cut, collected in a stomacher bag and rinsed with 250 mL buffered peptone water (BPW) (Oxoid, United Kingdom). For a first enrichment, 10 mL of this sample broth was added to 90 mL tryptic soy broth (TSB) (Oxoid, United Kingdom) with 6.5% NaCl and incubated at 37 °C overnight. Further, isolation was carried out as described previously [9] (link), with minor changes: 1 mL of the overnight culture was added to 5 mL VRE broth (Oxoid, United Kingdom) containing 16 µg/mL vancomycin. After shaking incubation at 42 °C overnight, one 10 µL-loop was spread on vancomycincontaining (16 µg/mL) Slanetz and Bartley agar (3% agar), which was incubated at 42 °C for 48 h. Two colonies per plate with typical enterococci morphology were transferred to Blood Agar Base plates (Oxoid, United Kingdom) (3% agar) and incubated at 37 °C overnight. Species identification was performed by MALDI-TOF MS. Enterococci strains were stored in 50% glycerol at -80 °C. Additional characterization was carried out with only one isolate per meat sample.
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