RESUMO
The pharmacokinetics of marbofloxacin in pigs after intravenous (i.v.), intramuscular (i.m.), and peroral (p.o.) administration and pharmacokinetic/pharmacodynamic indices of this drug against Korean local isolates of Actinobacillus pleuropneumoniae were determined in this study. Marbofloxacin (2.50 mg/kg of body weight) was administered, and blood samples were collected with designated time intervals. Plasma-extracted marbofloxacin was injected into the LC-MS/MS system. The in vitro and ex vivo antibacterial activities of marbofloxacin were evaluated against 20 isolates of A. pleuropneumoniae. The mean peak plasma concentrations (Cmax) after i.v., i.m., and p.o administration were 2.60 ± 0.10, 2.59 ± 0.12, and 2.34 ± 0.12 µg/mL at 0.25 ± 0.00, 0.44 ± 0.10, and 1.58 ± 0.40 h, respectively. The area under the plasma concentration-time curves (AUC0-24) and elimination half-lives were 24.80 ± 0.90, 25.80 ± 1.40, and 23.40 ± 5.00 h·µg/mL and 8.60 ± 0.30, 12.80 ± 1.10, and 8.60 ± 0.00 h, for i.v., i.m., and p.o. administration, correspondingly. The AUC0-24/MICs of marbofloxacin after i.v., i.m., and p.o. administration were 253.86 ± 179.91, 264.1 ± 187.16, and 239.53 ± 169.75 h, respectively. The Cmax/MIC values were 26.58 ± 18.84, 26.48 ± 18.77, and 23.94 ± 16.97, and T>MICs were 42.80 ± 1.01, 36.40 ± 1.24, and 38.60 ± 1.18 h, after i.v., i.m., and p.o. administration, respectively. Thus, marbofloxacin dosage of 2.50 mg/kg of body weight by i.v., i.m., and p.o. administration with 24 h dosing interval will provide effective treatment for the infection of pig by A. pleuropneumonia.
Assuntos
Infecções por Actinobacillus/tratamento farmacológico , Actinobacillus pleuropneumoniae , Fluoroquinolonas/farmacologia , Actinobacillus pleuropneumoniae/crescimento & desenvolvimento , Actinobacillus pleuropneumoniae/isolamento & purificação , Animais , Avaliação Pré-Clínica de Medicamentos , República da Coreia , SuínosRESUMO
BACKGROUND: Actinobacillus pleuropneumoniae is the etiologic agent of porcine contagious pleuropneumonia, which causes important worldwide economic losses in the swine industry. Several respiratory tract infections are associated with biofilm formation, and A. pleuropneumoniae has the ability to form biofilms in vitro. Biofilms are structured communities of bacterial cells enclosed in a self-produced polymer matrix that are attached to an abiotic or biotic surface. Virtually all bacteria can grow as a biofilm, and multi-species biofilms are the most common form of microbial growth in nature. The goal of this study was to determine the ability of A. pleuropneumoniae to form multi-species biofilms with other bacteria frequently founded in pig farms, in the absence of pyridine compounds (nicotinamide mononucleotide [NMN], nicotinamide riboside [NR] or nicotinamide adenine dinucleotide [NAD]) that are essential for the growth of A. pleuropneumoniae. RESULTS: For the biofilm assay, strain 719, a field isolate of A. pleuropneumoniae serovar 1, was mixed with swine isolates of Streptococcus suis, Bordetella bronchiseptica, Pasteurella multocida, Staphylococcus aureus or Escherichia coli, and deposited in 96-well microtiter plates. Based on the CFU results, A. pleuropneumoniae was able to grow with every species tested in the absence of pyridine compounds in the culture media. Interestingly, A. pleuropneumoniae was also able to form strong biofilms when mixed with S. suis, B. bronchiseptica or S. aureus. In the presence of E. coli, A. pleuropneumoniae only formed a weak biofilm. The live and dead populations, and the matrix composition of multi-species biofilms were also characterized using fluorescent markers and enzyme treatments. The results indicated that poly-N-acetyl-glucosamine remains the primary component responsible for the biofilm structure. CONCLUSIONS: In conclusion, A. pleuropneumoniae apparently is able to satisfy the requirement of pyridine compounds through of other swine pathogens by cross-feeding, which enables A. pleuropneumoniae to grow and form multi-species biofilms.
Assuntos
Infecções por Actinobacillus/veterinária , Actinobacillus pleuropneumoniae/crescimento & desenvolvimento , Actinobacillus pleuropneumoniae/metabolismo , Biofilmes/crescimento & desenvolvimento , NAD/deficiência , Acetilglucosamina/metabolismo , Infecções por Actinobacillus/microbiologia , Actinobacillus pleuropneumoniae/isolamento & purificação , Actinobacillus pleuropneumoniae/patogenicidade , Animais , Biofilmes/efeitos dos fármacos , Bordetella bronchiseptica/crescimento & desenvolvimento , Bordetella bronchiseptica/metabolismo , Meios de Cultura , Desoxirribonuclease I/farmacologia , Endopeptidase K/farmacologia , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/metabolismo , Hibridização in Situ Fluorescente , Microscopia Confocal , Niacinamida/análogos & derivados , Niacinamida/deficiência , Mononucleotídeo de Nicotinamida/deficiência , Pasteurella multocida/crescimento & desenvolvimento , Pasteurella multocida/metabolismo , Piridinas/metabolismo , Compostos de Piridínio , Especificidade da Espécie , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/metabolismo , Células-Tronco , Streptococcus suis/crescimento & desenvolvimento , Streptococcus suis/metabolismo , Suínos , Doenças dos Suínos/microbiologiaRESUMO
A real-time quantitative PCR (qPCR) for detection of the apxIVA gene of Actinobacillus pleuropneumoniae was validated using pure cultures of A. pleuropneumoniae and tonsillar and nasal swabs from experimentally inoculated Caesarean-derived/colostrum-deprived piglets and naturally infected conventional pigs. The analytical sensitivity was 5colony forming units/reaction. In comparison with selective bacterial examination using tonsillar samples from inoculated animals, the diagnostic sensitivity of the qPCR was 0.98 and the diagnostic specificity was 1.0. The qPCR showed consistent results in repeatedly sampled conventional pigs. Tonsillar brush samples and apxIVA qPCR analysis may be useful for further epidemiological studies and monitoring for A. pleuropneumoniae.
Assuntos
Infecções por Actinobacillus/veterinária , Actinobacillus pleuropneumoniae/genética , Proteínas de Bactérias/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Doenças dos Suínos/diagnóstico , Infecções por Actinobacillus/diagnóstico , Infecções por Actinobacillus/microbiologia , Actinobacillus pleuropneumoniae/isolamento & purificação , Animais , Cesárea/veterinária , Colostro/microbiologia , Nariz/microbiologia , Tonsila Palatina/microbiologia , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Sensibilidade e Especificidade , Análise de Sequência de DNA , Suínos , Doenças dos Suínos/microbiologiaRESUMO
SPF pigs aged 10 weeks were infected intranasally with Actinobacillus pleuropneumoniae serotype 2. After the onset of clinical symptoms of respiratory disease, which occurred 20 h post-infection, parenteral treatment with ceftiofur, danofloxacin, enrofloxacin, penicillin or tiamulin was initiated (n = 8 per group). Untreated groups, of which one was infected, served as controls. The uninfected control group did not show any signs of disease, while the infected control group was severely affected by the infection and also expressed a decreased weight gain following the challenge. Based on clinical signs, the magnitude of pathological lesions in the respiratory tract found at necropsy performed 17 days post-infection and the number of reisolates of A. pleuropneumoniae made at necropsy, treatments with the quinolones (danofloxacin and enrofloxacin) and the cephalosporine (ceftiofur) were superior to those with penicillin and tiamulin. The latter groups also developed antibodies to A. pleuropneumoniae to a larger extent. Some of the pigs treated with ceftiofur and danofloxacin developed antibodies to A. pleuropneumoniae, and the microbe was reisolated from approximately 50% of these animals. In contrast, pigs treated with enrofloxacin did not develop antibodies to A. pleuropneumoniae, and the challenge strain was not found at necropsy. The performance with respect to daily weight gain and feed conversion corresponded well with the clinical signs developed and the findings made at necropsy. The decreased growth recorded during the acute phase of the disease was, to a large extent, caused by a reduced feed intake.