Calmivet

For this prospective study, 40 healthy, non-castrated pet ferrets (Mustela putorius furo) were used. All 40 ferrets were subjects of the Clinical Veterinary Hospital of the University of Extremadura. They had no cardiorespiratory pathologies, as confirmed by physical examination, auscultation, and echocardiography, according to the reference values (25 (link), 26 (link)). An additional inclusion criterion was a minimum age of 6 months, since this is the age at which they reach their adult size and sexual maturity (27 ). Subjects who, after cardiac evaluation, suffered from some pathological process that could influence their cardiovascular status were excluded from the study.
In all ferrets, echocardiography and chest X-ray were performed under sedation [butorphanol (Torbugesic ® Vet, Zoetis, Madrid, Spain, 0.2 mg/kg), butorphanol-acepromazine (Torbugesic® Vet, Zoetis, Madrid, Spain, 0.2 mg/kg; Calmivet ®, Vetoquinol, Madrid, Spain, 0.1 mg/kg) or acepromazine-buprenorphine (Calmivet ®, Vetoquinol, Madrid, Spain, 0.1 mg/kg; Buprecare ®, Ecuphar NV, Oostkamp, Belgium, 0.01 mg/kg)], due to the low cardiovascular effects. Data such as age, sex, and body weight were routinely collected.

Thirty minutes after removal from the hyperbaric chamber, the rats were anaesthetised with an intraperitoneal injection of a mixture of 10 mg.kg⁻¹ xylazine (Rompum® 2%, Bayer Pharma, Germany), 100 mg.kg⁻¹ ketamine (Imalgène®1000, Merial, France) and 1.65 mg.kg⁻¹ acepromazine (Calmivet®, Vetoquinol, France).
At the end of the study, the rats were sacrificed by means of an intraperitoneal injection of pentobarbital (200 mg.kg⁻¹, Sanofi Santé, France).

BALB/cJ mice were purchased from Janvier (Le Genest St. Isle, France). Mice were housed under standard conditions of feeding, light and temperature with access to food and water ad libidium. Experiments were performed according to the national and Institut Pasteur guidelines for laboratory animal experiments. Protocols were approved by the Institut Pasteur animal care and use committee (protocol 05–59) and the Direction des Services Vétérinaire de Paris (permit 75–713 to RT). Six- to eight-week-old mice were anesthetized with acepromazine (Calmivet, 1.5 mg/kg, Vetoquinol, Lure, France). and ketamine (Imalgene, 31.25 mg/kg, Merial, Lyon, France). and then infected intranasally with 20 μl bacterial suspension. The virulence of K. pneumoniae strains was tested on six-week-old BALB/c mice, as previously described [23 (link)]. Seven mice per test condition were infected with 10⁸ bacteria. Mice were followed every day for one week. Experiments were performed at least twice.