Pharmacokinetic profile and effect on the faecal microbiome of a single dose of pradofloxacin oral suspension in the rabbit (Oryctolagus cuniculus).

Fluoroquinolones are often administered to pet rabbits given their perceived safety and limited effects on anaerobic gut microbiota. However, the pharmacokinetics and relative safety of pradofloxacin, a third-generation veterinary fluoroquinolone with a much broader spectrum of activity, have not been reported in this species. Here, we determined the pharmacokinetic profile of a single dose of oral pradofloxacin in rabbits and evaluated effects on the faecal microbiome. Four mature female rabbits were administered pradofloxacin (25 mg/ml oral suspension), at a dose of 7.5 mg/kg. The pradofloxacin median (range) Tmax was 4.50 (2.00-5.00) h, Cmax 600.66 (395.85-886.72) ng/ml and t½ was 1.27 (0.12-1.39) h. These results indicated that oral absorption of pradofloxacin was slower, and elimination faster compared with other fluoroquinolones in healthy rabbits, as well as relative to cats and dogs. Following treatment with pradofloxacin, faecal microbiota profiling showed some compositional differences between treated and control animals. This was the result of a significant decrease in the abundance of Proteobacteria, in particular bacteria belonging to the Pseudomonas, Atopostipes and Parabacteroides genera. The pharmacokinetic profile of pradofloxacin in rabbits should be further studied by increasing the sample size and using multiple-dose protocols (i.e. 7 days) to confirm safety. Further information on the effects of protein binding, higher dosages and disease on pradofloxacin pharmacokinetics in rabbits are needed before an accurate dosing regimen can be recommended.

Pharmacokinetic profile of a single dose of an oral pradofloxacin suspension administered to eastern long-necked turtles (Chelodina longicollis).

The pharmacokinetics of fluoroquinolones in chelonians are well described but this does not extend to pradofloxacin, a broad-spectrum veterinary fluoroquinolone available as an oral suspension for cats and dogs. The aim of this study was to investigate the single-dose pharmacokinetic profile of pradofloxacin oral suspension at 7.5 mg/kg in eastern long-necked turtles (Chelodina longicollis). Eight treated turtles were sampled at multiple time points up to 168 hr. Plasma concentrations were measured using high-performance liquid chromatography. Pradofloxacin was quantifiable for up to 48 hr after drug administration. The Tmax (9.0 hr) and T½ to 48 hr (13.16 hr) were longer, and the Cmax (0.2 µg/ml) and AUC0-24 (2.2 hr*µg/ml) lower, than previously reported in cats and dogs. Pradofloxacin was measurable in tank water samples for up to 48 hr. No adverse effects were observed in six turtles administered 7.5 mg/kg sid for 7 days. Using mammalian MIC data, the AUC0-24 /MIC ratios for a range of bacterial isolates suggest that this dose of pradofloxacin in turtles is unlikely to be effective against many bacterial pathogens.

Influence of long-term oral application of quinolones on the ECG curve in dogs.

The aim of the study was to analyse the influence of enrofloxacin and pradofloxacin administered orally for 14 days on the ECG in dogs. The ECG was performed before and after a 14 day period of quinolone administration. There was an increase in the QTc and the TpTe interval in the group treated with quinolones. QTc was prolonged by 24 ms (p=0.001). The TpTe interval was shortened, on average, by 6.55 ms (p=0.048). In the group treated with enrofloxacin, QTc was prolonged by 16.27 ms (p=0.006) and the TpTe interval was shortened by 9.64 ms (p=0.050), the TpTe/QT index was reduced by 0.034 (p=0.050) on average. In dogs treated with pradofloxacin, QTc was prolonged by 21.55 ms (p=0.012) on average. The results suggest that a prolonged administration of quinolones can increase the risk of arrhythmias. Furthermore, different generations of these drugs increase this risk to various degrees. The study proved that second generation quinolones, such as enrofloxacin, significantly change the phase of depolarization and repolarization of the ventricles, at the same time increasing the risk of ventricular arrythmia. Pradofloxacin does not change the TpTe and TpTe/QT values, so it is safer in use.

Pradofloxacin for Treatment of Bartonella henselae in Experimentally Inoculated Cats.

Bartonella henselae is associated with numerous clinical syndromes in people. Cats are the definitive hosts for B. henselae, develop high levels of bacteremia, and are associated with human infections, particularly in the presence of Ctenocephalides felis. Several antibiotic protocols used for the treatment of B. henselae infection in cats have failed to clear bacteremia. The purpose of this study was to assess the safety and efficacy of a high-dose pradofloxacin protocol to eliminate B. henselae bacteremia. Bartonella henselae infection was initiated in 8 cats by intravenous inoculation of infected feline blood and then pradofloxacin was administered at 7.5 mg/kg, PO, twice daily for 28 days, starting 12 weeks after inoculation. Complete blood cell counts were performed prior to pradofloxacin administration and then every 2 weeks for 10 weeks. Bartonella PCR assay was performed prior to pradofloxacin administration and approximately every 2 weeks for 10 weeks and then weekly for 4 weeks. Methylprednisolone acetate (5 mg/kg) was administered by intramuscular injection to all cats on week 10. The cats remained normal and none developed a hematocrit, platelet count, lymphocyte count, or neutrophil count outside of the normal reference ranges. In the one month prior to pradofloxacin administration, all cats were PCR-positive for Bartonella DNA on at least two of four sample dates; after pradofloxacin administration, all cats were negative for B. henselae DNA in blood on all nine sample dates. The protocol appears to be safe and failure to amplify B. henselae DNA from the blood after the administration of pradofloxacin and one dose of methylprednisolone acetate suggests either an antibiotic effect or the organism was cleared spontaneously.

Comparative In Vitro Killing by Pradofloxacin in Comparison to Ceftiofur, Enrofloxacin, Florfenicol, Marbofloxacin, Tildipirosin, Tilmicosin and Tulathromycin against Bovine Respiratory Bacterial Pathogens.

Pradofloxacin is the newest of the veterinary fluoroquinolones to be approved for use in animals-initially companion animals and most recently food animals. It has a broad spectrum of in vitro activity, working actively against Gram-positive/negative, atypical and some anaerobic microorganisms. It simultaneously targets DNA gyrase (topoisomerase type II) and topoisomerase type IV, suggesting a lower propensity to select for antimicrobial resistance. The purpose of this study was to determine the rate and extent of bacterial killing by pradofloxacin against bovine strains of Mannheimia haemolytica and Pasteurella multocida, in comparison with several other agents (ceftiofur, enrofloxacin, florfenicol, marbofloxacin, tildipirosin, tilmicosin and tulathromycin) using four clinically relevant drug concentrations: minimum inhibitory and mutant prevention drug concentration, maximum serum and maximum tissue drug concentrations. At the maximum serum and tissue drug concentrations, pradofloxacin killed 99.99% of M. haemolytica cells following 5 min of drug exposure (versus growth to 76% kill rate for the other agents) and 94.1-98.6% of P. multocida following 60-120 min of drug exposure (versus growth to 98.6% kill rate for the other agents). Statistically significant differences in kill rates were seen between the various drugs tested depending on drug concentration and time of sampling after drug exposure.

Comparative Minimum Inhibitory and Mutant Prevention Drug Concentrations for Pradofloxacin and Seven Other Antimicrobial Agents Tested against Bovine Isolates of Mannheimia haemolytica and Pasteurella multocida.

Pradofloxacin-a dual-targeting fluoroquinolone-is the most recent approved for use in food animals. Minimum inhibitory and mutant prevention concentration values were determined for pradofloxacin, ceftiofur, enrofloxacin, florfenicol, marbofloxacin, tildipirosin, tilmicosin, and tulathromycin. For M. haemolytica strains, MIC50/90/100 values were ≤0.016/≤0.016/≤0.016 and MPC50/90/100 values were 0.031/0.063/0.063; for P. multocida strains, the MIC50/90/100 values ≤0.016/≤0.016/0.031 and MPC50/90/100 ≤ 0.016/0.031/0.063 for pradofloxacin. The pradofloxacin Cmax/MIC90 and Cmax/MPC90 values for M. haemolytica and P. multocida strains, respectively, were 212.5 and 53.9 and 212.5 and 109.7. Similarly, AUC24/MIC90 and AUC24/MPC90 for M. haemolytica were 825 and 209.5, and for P. multocida, they were 825 and 425.8. Pradofloxacin would exceed the mutant selection window for >12-16 h. Pradofloxacin appears to have a low likelihood for resistance selection against key bovine respiratory disease bacterial pathogens based on low MIC and MPC values.

Treatment of a cat with presumed Bartonella henselae-associated immune-mediated hemolytic anemia, fever, and lymphadenitis.

A 2.5-year-old castrated male cat presented with fever and marked generalized lymphadenopathy of 4-months duration, despite treatment with amoxicillin-clavulanate/marbofloxacin. Abnormalities were not detected on complete blood count, serum chemistry, and FIV/FeLV test apart from a borderline, non-regenerative anemia. Peripheral lymph node fine needle aspirations revealed a marked increase in the percentage of intermediate- and lymphoblastic-lymphocytes in addition to reactive macrophages. Three weeks after presentation, the cat developed a severe, regenerative, immune-mediated hemolytic anemia (IMHA) which responded to immunosuppressive therapy. Fever and lymphadenopathy persisted. Peripheral lymph nodes tested positive for Bartonella henselae DNA in real-time PCR assay and sequencing. Treatment with pradofloxacin and doxycycline resulted in resolution of clinical signs, and negative PCR tests. Despite its reported low pathogenicity, B. henselae infection should also be considered in cats with protracted unexplained fever, lymphadenitis, and IMHA. Furthermore, a combination of pradofloxacin and doxycycline might be considered in cats with bartonellosis given its apparent clinical efficacy.

Cutaneous Mycobacterium goodii infection in an immunocompetent cat in Louisiana: clinical presentation, molecular identification, antimicrobial susceptibility and management.

Case summary: A 9-year-old spayed female domestic shorthair cat was presented to a referral hospital for management of recurring non-healing ulcerations and a subcutaneous mass on the ventral abdomen. Prior treatment included antibiotics (cefovecin followed by clindamycin), wound cleaning and surgical debulking, but the ulcerations and mass recurred 1 month after surgical removal. At this point, the cat was started on doxycycline and pradofloxacin and referred for further work-up. The culture of skin biopsy specimens obtained at the time of referral revealed a population of bacterial colonies with two distinctly different phenotypes. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and 16S rRNA gene sequencing identified both colonies as Mycobacterium goodii. A diagnosis of a cutaneous infection of rapidly growing mycobacteria was made, and treatment with oral pradofloxacin and doxycycline was initiated. The ulcerations resolved within 4 months, and the subcutaneous mass gradually decreased in size until it was no longer palpable, even 4 months after the cessation of antibiotics. Relevance and novel information: This is the second reported feline cutaneous M goodii infection in North America. The organism was not visualized on histopathology but was successfully cultured from tissue obtained by skin punch biopsy. A phenotypic switching phenomenon affecting the susceptibility results was suspected, possibly explaining the presence of phenotypically different but genetically identical strains. This case highlights the importance of submitting aseptically obtained tissue, fluid or fine-needle aspirates for culture and species identification, as well as histopathology, when infection with higher bacteria, such as rapidly growing mycobacteria, is suspected.

Resistance of canine methicillin-resistant Staphylococcus pseudintermedius strains to pradofloxacin.

We investigated in vitro activity of a novel veterinary fluoroquinolone, pradofloxacin, against methicillin-resistant Staphylococcus pseudintermedius (MRSP) isolates and compared with other fluoroquinolones. A total of 38 MRSP isolates were subjected to agar disk diffusion tests for sensitivity to pradofloxacin, orbifloxacin, marbofloxacin, enrofloxacin, and ciprofloxacin. The minimal inhibitory concentration (MIC) values of pradofloxacin, ciprofloxacin, and enrofloxacin were determined. Mutations in the genes encoding DNA gyrase subunit A (GyrA) and topoisomerase IV (GrlA) proteins associated with fluoroquinolone resistance were studied by an analysis of partial sequences of the genes encoding these proteins. Two MRSP isolates were susceptible in disk diffusion and microdilution test to all fluoroquinolones tested, including pradofloxacin. Based on the results of the disk diffusion testing, 33 of 38 isolates showed resistance to pradofloxacin and 3 were intermediate, whereas, by pradofloxacin MIC testing, 35 isolates were classified as resistant and 1 as intermediate. Single alterations in GyrA and GrlA proteins were observed in the 35 resistant isolates and the 1 intermediate isolate (MIC results). These same 36 isolates were also resistant to the other tested fluoroquinolones. The results of the current study showed that MRSP isolates are usually resistant to all fluoroquinolones, including pradofloxacin. Therefore, in routine susceptibility testing to pradofloxacin by disk diffusion, the results should be carefully interpreted for MRSP isolates, especially those resistant to other fluoroquinolones and, in questionable cases, the pradofloxacin MIC should be determined to confirm the susceptibility testing results.

In vitro selection of resistance to pradofloxacin and ciprofloxacin in canine uropathogenic Escherichia coli isolates.

This study explored and compared the mechanisms and selective concentration of resistance between a 3rd (pradofloxacin) and 2nd (ciprofloxacin) generation fluoroquinolone. Pradofloxacin- and ciprofloxacin-resistant mutants were selected by stepwise exposure of Escherichia coli (E. coli) to escalating concentrations of pradofloxacin and ciprofloxacin. The sequence of the quinolone resistance determining region (QRDR) and the transcriptional regulator soxS were analyzed, and efflux pump AcrAB-TolC activity was measured by quantitative real-time reverse transcription-PCR (qRT-PCR). First-step mutants reduced the fluoroquinolone sensitivity and one mutant bore a single substitution in gyrA. Four of six second-step mutants expressed ciprofloxacin resistance, and displayed additional mutations in gyrA and/or parC, while these mutants retained susceptibility to pradofloxacin. All the third-step mutants were fluoroquinolone resistant, and each expressed multidrug resistance (MDR) phenotypes. Further, they displayed resistance to all antibacterials tested except cefotaxime, ceftazidime and meropenem. The number of mutations in QRDR of gyrA and parC correlated with fluoroquinolone MICs. Mutations in parC were not common in pradofloxacin-associated mutants. Moreover, one second- and one third-step ciprofloxacin-associated mutants bore both mutations at position 12 (Ala12Ser) and 78 (Met78Leu) in the soxS gene, yet no mutations in the soxS gene were detected in the pradofloxacin-selected mutants. Altogether, these results demonstrated that resistance emerged relatively more rapidly in 2nd compared to 3rd generation fluoroquinolones. Point mutations in gyrA were a key mechanism of resistance to pradofloxacin, and overexpression of efflux pump gene acrB played a potential role in the emergence of MDR phenotypes identified in this study.