Table 3.
Groups of animals used in animal study.
| Group | Purpose of experiment | Number of animals | UTI induction/used strain | Administrated compound | Dose of administrated compound | Urine culture | Route of drug administration |
|---|---|---|---|---|---|---|---|
| 1 | Control | 5 | – | 0,9% sterile saline | 0.1 mL | + | Intravenously |
| 2 | Antibacterial efficiency | 5 | E. coli Xen14 | 0,9% sterile saline | 0.1 mL | + | |
| 3 | 5 | E. coli Xen14 | CSA-13-IRDye 800CW | 10 µg/mL (including 1 µg/mL of IRDye® 800CW) | + | ||
| 4 | 5 | E. coli clinical strain (S1) | 0,9% sterile saline | 0.1 mL | + | ||
| 5 | 5 | E. coli clinical strain (S1) | CSA-13 | 10 µg/mL | + | ||
| 6 | 5 | E. coli clinical strain (S2) | 0,9% sterile saline | 0.1 mL | + | ||
| 7 | 5 | E. coli clinical strain (S2) | CSA-13 | 10 µg/mL | + | ||
| 8 | Biodistibution assesment | 5 | – | CSA-13-IRDye 800CW | 10 µg/mL (including 1 µg/mL of IRDye® 800CW) | − | Intravenously |
| 9 | 5 | – | IRDye 800CW | 1 µg/mL | − |
Urinary tract infection (Day 0) induction of urinary tract infection by E. coli Xen14 and two clinical strain of E. coli (S1 and S2) administration (n = 5 per group; group 2–7). (Day 1) 24 h after induction, mice were divided into 2 groups: (1) UTI-suffering, untreated animals that obtained intravenously 0.1 mL of 0.9% NaCl (n = 5; group 2, 4, 6), and (2) UTI-suffering animals, treated i.v. with CSA-13-IRDye800CW or CSA-13 at dosage of 5 µg/g of body weight (n = 5 per each group; group 3,5,7). (Day 2) the urine was collected from the animals from 1 to 7 groups at the specified times (0 h, 4 h, 8 h, 12 h and 24 h after treatment) and then were plated on MacConkey agar plates to determine bacterial titers.
Part 2: Biodistribution of CSA-13. (Day 0) Mice were injected with 0.1 mL of 0.9% NaCl (n = 15). (Day 1) mice were divided into the following groups: Group 8 and 9 were injected intravenously with CSA-13-IRDye800CW (n = 5; group 8) or unmodified IRDye800CW dye (n = 5; group 9), respectively. (Day 2) Animals from 8 and 9 groups were imaged using the Pearl®Trilogy small animal imaging system (Li-COR, Lincoln, NB, USA) at different times after treatment with CSA-13-IRDye800CW (n = 5; group 8) or unmodified IRDye800CW dye (n = 5; group 9) (0 h, 4 h, 8 h, and 12 h), after that animals from all groups were sacrificed and tissues were analyzed.