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The spread of antibiotic resistance is one of the biggest challenges of our time, making it difficult to treat bacterial diseases. Pasteurella multocida is a widespread facultative pathogenic bacterium, which causes a wide range of diseases in both mammals and birds. In the present study, antibiotic susceptibility of 155 P. multocida strains were tested using the broth microdilution method to obtain the minimum inhibitory concentration (MIC) values for 15 antibiotics. The most effective antibiotics against pasteurellosis were ceftiofur, tetracycline, doxycycline, florfenicol and tilmicosin. Of the strains, 12 proved to be multi-drug resistant (MDR). To combat antibiotic resistance, it is important to establish a pre-treatment antibiotic susceptibility profile. A well-chosen antibiotic would not only make the treatment more successful but may also slow down the spread of resistance and the evolution of MDR strains.
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Antiinfecciosos , Farmacorresistencia Bacteriana , Pasteurella multocida , Pasteurella multocida/efectos de los fármacos , Pasteurella multocida/aislamiento & purificación , Antiinfecciosos/farmacología , Pruebas de Sensibilidad Microbiana , Infecciones por Pasteurella/microbiología , Farmacorresistencia Bacteriana Múltiple , Aves/microbiología , Mamíferos/microbiología , Animales , BovinosRESUMEN
Lv17/WB/Rie1-Δ24 was produced via illegitimate recombination mediated by low-dilution serial passage in the Cos7 cell line and isolated on PAM cell culture. The virus contains a huge ~26.4 Kb deletion in the left end of its genome. Lv17/WB/Rie1-ΔCD-ΔGL was generated via homologous recombination, crossing two ASFV strains (Lv17/WB/Rie1-ΔCD and Lv17/WB/Rie1-ΔGL containing eGFP and mCherry markers) during PAM co-infection. The presence of unique parental markers in the Lv17/WB/Rie1-ΔCD-ΔGL genome indicates at least two recombination events during the crossing, suggesting that homologous recombination is a relatively frequent event in the ASFV genome during replication in PAM. Pigs infected with Lv17/WB/Rie1-Δ24 and Lv17/WB/Rie1/ΔCD-ΔGL strains have shown mild clinical signs despite that ASFV could not be detected in their sera until a challenge infection with the Armenia/07 ASFV strain. The two viruses were not able to induce protective immunity in pigs against a virulent Armenia/07 challenge.
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African swine fever (ASF) is a highly lethal hemorrhagic viral disease that causes extensive economic and animal welfare losses in the Eurasian pig (Sus scrofa) population. To date, no effective and safe vaccines have been marketed against ASF. A starting point for vaccine development is using naturally occurring attenuated strains as a vaccine base. Here, we aimed to remove the multigene family (MGF) 110 gene of unknown function from the Lv17/WB/Rie1 genome to improve the usability of the virus as a live-attenuated vaccine, reducing unwanted side effects. The MGF 110-11L gene was deleted using the CRISPR/Cas9 method, and the safety and efficacy of the virus were tested in pigs after isolation. The vaccine candidates administered at high doses showed reduced pathogenicity compared to the parental strain and induced immunity in vaccinated animals, although several mild clinical signs were observed. Although Lv17/WB/Rie1/d110-11L cannot be used as a vaccine in its current form, it was encouraging that the undesirable side effects of Lv17/WB/Rie1 at high doses can be reduced by additional mutations without a significant reduction in its protective capacity.
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Vagal nerve stimulation (VNS) holds a strong basis as a potentially effective treatment modality for chronic heart failure, which explains why a multicenter VNS study in heart failure with reduced ejection fraction is ongoing. However, more detailed information is required on the effect of acetylcholine (ACh) on repolarization in Purkinje and ventricular cardiac preparations to identify the advantages, risks, and underlying cellular mechanisms of VNS. Here, we studied the effect of ACh on the action potential (AP) of canine Purkinje fibers (PFs) and several human ventricular preparations. In addition, we characterized the effects of ACh on the L-type Ca2+ current (ICaL) and AP of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) and performed computer simulations to explain the observed effects. Using microelectrode recordings, we found a small but significant AP prolongation in canine PFs. In the human myocardium, ACh slightly prolonged the AP in the midmyocardium but resulted in minor AP shortening in subepicardial tissue. Perforated patch-clamp experiments on hiPSC-CMs demonstrated that 5 µM ACh caused an ≈15% decrease in ICaL density without changes in gating properties. Using dynamic clamp, we found that under blocked K+ currents, 5 µM ACh resulted in an ≈23% decrease in AP duration at 90% of repolarization in hiPSC-CMs. Computer simulations using the O'Hara-Rudy human ventricular cell model revealed that the overall effect of ACh on AP duration is a tight interplay between the ACh-induced reduction in ICaL and ACh-induced changes in K+ currents. In conclusion, ACh results in minor changes in AP repolarization and duration of canine PFs and human ventricular myocardium due to the concomitant inhibition of inward ICaL and outward K+ currents, which limits changes in net repolarizing current and thus prevents major changes in AP repolarization.
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Macrolides are commonly used to control respiratory tract infections in ruminants, but the susceptibility of Pasteurella multocida strains has shown a decrease to macrolide antibiotics in the last decade. In this work we assessed the prevalence of macrolide resistance of 100 P. multocida isolates from ruminant hosts and studied the resistance genotypes with newly designed PCRs. Susceptibility to erythromycin and tilmicosin was tested using minimal inhibitory concentration (MIC) test strips. A newly designed PCR was used for the detection of macAB genes, and a PCR plus restriction enzyme-based technique was developed for detecting a 23S rRNA gene mutation at position 2059. Five bovine isolates with notably increased MICs (≥256 µg/mL for erythromycin and ≥32 µg/mL for tilmicosin) carried resistance genes msr(E) and mph(E) or the A2059G point mutation in the 23S rRNA gene. Over 73% strains from small ruminants and all bovine isolates were MacAB PCR positive. Bovine strains were less sensitive to macrolide antibiotics than isolates from small ruminants, and an increase in the prevalence of macrolide resistance in bovine P. multocida isolates has also been observed over time.
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Pasteurella multocida , Animales , Antibacterianos/farmacología , Bovinos , Farmacorresistencia Bacteriana/genética , Eritromicina , Genotipo , Macrólidos/farmacología , Pruebas de Sensibilidad Microbiana , Pasteurella multocida/genética , ARN Ribosómico 23S/genética , Rumiantes/genética , Tilosina/análogos & derivadosRESUMEN
Vagal nerve stimulation (VNS) has a meaningful basis as a potentially effective treatment for heart failure with reduced ejection fraction. There is an ongoing VNS randomized study, and four studies are completed. However, relatively little is known about the effect of acetylcholine (ACh) on repolarization in human ventricular cardiomyocytes, as well as the effect of ACh on the rapid component of the delayed rectifier K+ current (IKr). Here, we investigated the effect of ACh on the action potential parameters in human ventricular preparations and on IKr in human induced pluripotent stem-cell-derived cardiomyocytes (hiPSC-CMs). Using standard microelectrode technique, we demonstrated that ACh (5 µM) significantly increased the action potential duration in human left ventricular myocardial slices. ACh (5 µM) also prolonged repolarization in a human Purkinje fiber and a papillary muscle. Optical mapping revealed that ACh increased the action potential duration in human left ventricular myocardial slices and that the effect was dose-dependent. Perforated patch clamp experiments demonstrated action potential prolongation and a significant decrease in IKr by ACh (5 µM) in hiPSC-CMs. Computer simulations of the electrical activity of a human ventricular cardiomyocyte showed an increase in action potential duration upon implementation of the experimentally observed ACh-induced changes in the fully activated conductance and steady-state activation of IKr. Our findings support the hypothesis that ACh can influence the repolarization in human ventricular cardiomyocytes by at least changes in IKr.
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BACKGROUND AND PURPOSE: The aim of the present study was to study the antiarrhythmic effects and cellular mechanisms of desethylamiodarone (DEA), the main metabolite of amiodarone (AMIO), following acute and chronic 4-week oral treatments (25-50 mg·kg-1 ·day-1 ). EXPERIMENTAL APPROACH: The antiarrhythmic effects of acute iv. (10 mg·kg-1 ) and chronic oral (4 weeks, 25 mg·kg-1 ·day-1 ) administration of DEA were assessed in carbachol and tachypacing-induced dog atrial fibrillation models. Action potentials were recorded from atrial and right ventricular tissue following acute (10 µM) and chronic (p.o. 4 weeks, 50 mg·kg-1 ·day-1 ) DEA application using the conventional microelectrode technique. Ionic currents were measured by the whole cell configuration of the patch clamp technique in isolated left ventricular myocytes. Pharmacokinetic studies were performed following a single intravenous dose (25 mg·kg-1 ) of AMIO and DEA intravenously and orally. In chronic (91-day) toxicological investigations, DEA and AMIO were administered in the oral dose of 25 mg·kg-1 ·day-1 ). KEY RESULTS: DEA exerted marked antiarrhythmic effects in both canine atrial fibrillation models. Both acute and chronic DEA administration prolonged action potential duration in atrial and ventricular muscle without any changes detected in Purkinje fibres. DEA decreased the amplitude of several outward potassium currents such as IKr , IKs , IK1 , Ito , and IKACh , while the ICaL and late INa inward currents were also significantly depressed. Better drug bioavailability and higher volume of distribution for DEA were observed compared to AMIO. No neutropenia and less severe pulmonary fibrosis was found following DEA compared to that of AMIO administration. CONCLUSION AND IMPLICATIONS: Chronic DEA treatment in animal experiments has marked antiarrhythmic and electrophysiological effects with better pharmacokinetics and lower toxicity than its parent compound. These results suggest that the active metabolite, DEA, should be considered for clinical trials as a possible new, more favourable option for the treatment of cardiac arrhythmias including atrial fibrillation.
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Amiodarona , Fibrilación Atrial , Potenciales de Acción , Amiodarona/análogos & derivados , Amiodarona/farmacología , Animales , Antiarrítmicos/farmacología , Fibrilación Atrial/tratamiento farmacológico , Fibrilación Atrial/metabolismo , Perros , Atrios Cardíacos , Miocitos CardíacosRESUMEN
The ability to distinguish among the subspecies of Pasteurella multocida isolates is important epidemiologically; however, classification at the subspecies level based on the results of conventional biochemical tests (fermentation of sorbitol and dulcitol) is reportedly not accurate in all cases. Therefore, we developed a rapid, multiplex PCR assay to differentiate among the 3 subspecies of P. multocida. The PCR assay includes the P. multocida species-specific primers KMT1SP6 and KMT1T7 as an internal amplification control, with a newly designed gatD (galactitol-1-phosphate-5-dehydrogenase)-specific primer pair (unique for subsp. gallicida), and primers targeting a 16S rRNA gene region specific for subsp. septica. The subspecies specificity of the PCR was demonstrated by applying the test to a collection of 70 P. multocida isolates, including the Heddleston serovar reference strains; all isolates and strains were assigned correctly. The PCR assay is a sensitive, specific, and highly effective method for the identification of P. multocida subspecies, and an alternative to biochemical test-based differentiation. A possible relationship was noticed between P. multocida subspecies and lipopolysaccharide (LPS) genotype; all but one of the subsp. gallicida strains were isolated only from avian hosts and represented L1 LPS genotype. Subsp. multocida and subsp. septica isolates were classified into 5 and 4 different LPS genotypes, respectively, of which L3 was the only LPS genotype shared between these 2 subspecies.
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Infecciones por Pasteurella , Pasteurella multocida , Animales , Genotipo , Reacción en Cadena de la Polimerasa Multiplex/veterinaria , Infecciones por Pasteurella/diagnóstico , Infecciones por Pasteurella/veterinaria , Pasteurella multocida/genética , ARN Ribosómico 16S/genéticaRESUMEN
Since 2010, outbreaks of haemorrhagic septicaemia (HS) caused by Pasteurella (P.) multocida capsular type B (PmB) emerged in Germany. In 2017, we noticed a close spatiotemporal relationship between HS outbreak sites and wolf (Canis lupus) territories. Thus, the main objectives of our study were to investigate the molecular epidemiology of German PmB-HS-isolates and to assess the role of wolves as putative vectors of this pathogen. We collected 83 PmB isolates from HS outbreaks that occurred between 2010 and 2019 and sampled 150 wolves, which were found dead in the years 2017 to 2019, revealing another three PmB isolates. A maximum-likelihood-based phylogeny of the core genomes of 65 PmB-HS-isolates and the three PmB-wolf-isolates showed high relatedness. Furthermore, all belonged to capsular:LPS:MLST genotype B:L2:ST122RIRDC and showed highly similar virulence gene profiles, but clustered separately from 35 global ST122RIRDC strains. Our data revealed that German HS outbreaks were caused by a distinct genomic lineage of PmB-ST122 strains, hinting towards an independent, ongoing epidemiologic event. We demonstrated for the first time, that carnivores, i.e., wolves, might harbour PmB as a part of their oropharyngeal microbiota. Furthermore, the results of our study imply that wolves can carry the pathogen over long distances, indicating a major role of that animal species in the ongoing epidemiological event of HS in Germany.
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Ibuprofen is a widely used nonsteroidal anti-inflammatory drug, which has recently been associated with increased cardiovascular risk, but its electrophysiological effects have not yet been properly studied in isolated cardiac preparations. We studied the effects of ibuprofen on action potential characteristics and several transmembrane ionic currents using the conventional microelectrode technique and the whole-cell configuration of the patch-clamp technique on cardiac preparations and enzymatically isolated ventricular myocytes. In dog (200 µM; n = 6) and rabbit (100 µM; n = 7) papillary muscles, ibuprofen moderately but significantly prolonged repolarization at 1 Hz stimulation frequency. In dog Purkinje fibers, repolarization was abbreviated and maximal rate of depolarization was depressed in a frequency-dependent manner. Levofloxacin (40 µM) alone did not alter repolarization, but augmented the ibuprofen-evoked repolarization lengthening in rabbit preparations (n = 7). In dog myocytes, ibuprofen (250 µM) did not significantly influence IK1, but decreased the amplitude of Ito and IKr potassium currents by 28.2% (60 mV) and 15.2% (20 mV), respectively. Ibuprofen also depressed INaL and ICa currents by 19.9% and 16.4%, respectively. We conclude that ibuprofen seems to be free from effects on action potential parameters at lower concentrations. However, at higher concentrations it may alter repolarization reserve, contributing to the observed proarrhythmic risk in patients.
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Potenciales de Acción/efectos de los fármacos , Antiinflamatorios no Esteroideos/efectos adversos , Arritmias Cardíacas/diagnóstico , Ventrículos Cardíacos/efectos de los fármacos , Ibuprofeno/efectos adversos , Animales , Antiinflamatorios no Esteroideos/administración & dosificación , Arritmias Cardíacas/inducido químicamente , Perros , Relación Dosis-Respuesta a Droga , Ventrículos Cardíacos/citología , Humanos , Ibuprofeno/administración & dosificación , Masculino , Microelectrodos , Miocitos Cardíacos , Técnicas de Placa-Clamp , Ramos Subendocárdicos/efectos de los fármacos , ConejosRESUMEN
Activation of the parasympathetic nervous system has been reported to have an antiarrhythmic role during ischemia-reperfusion injury by decreasing the arrhythmia triggers. Furthermore, it was reported that the parasympathetic neurotransmitter acetylcholine is able to modulate the ATP-dependent potassium current (I K-ATP), a crucial current activated during hypoxia. However, the possible significance of this current modulation in the antiarrhythmic mechanism is not fully clarified. Action potentials were measured using the conventional microelectrode technique from canine left ventricular papillary muscle and free-running Purkinje fibers, under normal and hypoxic conditions. Ionic currents were measured using the whole-cell configuration of the patch-clamp method. Acetylcholine at 5 µmol/L did not influence the action potential duration (APD) either in Purkinje fibers or in papillary muscle preparations. In contrast, it significantly lengthened the APD and suppressed the Purkinje-ventricle APD dispersion when it was administered after 5 µmol/L pinacidil application. Carbachol at 3 µmol/L reduced the pinacidil-activated I K-ATP under voltage-clamp conditions. Acetylcholine lengthened the ventricular action potential under simulated ischemia condition. In this study, we found that acetylcholine inhibits the I K-ATP and thus suppresses the ventricle-Purkinje APD dispersion. We conclude that parasympathetic tone may reduce the arrhythmogenic substrate exerting a complex antiarrhythmic mechanism during hypoxic conditions.
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Potenciales de Acción/efectos de los fármacos , Adenosina Trifosfato/metabolismo , Ventrículos Cardíacos/efectos de los fármacos , Agonistas Muscarínicos/farmacología , Potasio/metabolismo , Ramos Subendocárdicos/efectos de los fármacos , Animales , Perros , Ventrículos Cardíacos/citología , Ramos Subendocárdicos/citologíaRESUMEN
The occurrence of members of the Pasteurellaceae and Neisseriaceae families was studied in dogs and cats. A total of 110 nasal and pharyngeal swab samples from 47 dogs and 8 cats were collected. Most of the strains were identified by 16S rDNA sequencing, except Frederiksenia canicola and Pasteurella multocida where species-specific polymerase chain reactions were applied. The most frequently isolated species was F. canicola, which occurred only in dogs, mainly in the pharyngeal cavity. The second commonest bacterium, P. multocida was found in both types of samples and in both hosts. Other species from the family Pasteurellaceae, such as Haemophilus haemoglobinophilus, Pasteurella canis and P. dagmatis, were detected only in dogs. All isolated species belonging to the family Neisseriaceae, mainly representing Neisseria weaveri, were found only in the pharyngeal cavity. Neisseria weaveri and N. zoodegmatis could be detected in both hosts. Neisseria dumasiana and N. canis were isolated from dogs, while N. shayeganii only from a cat. For phylogenetic analysis, rpoB gene sequencing was performed, where the strains were on monophyletic branches and clearly separated from each other. In this study, recently described species such as F. canicola, N. shayeganii and N. dumasiana were detected that had never been isolated in Hungary before.
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Enfermedades de los Gatos/epidemiología , Enfermedades de los Perros/epidemiología , Infecciones por Bacterias Gramnegativas/veterinaria , Microbiota , Enfermedades Respiratorias/veterinaria , Animales , Enfermedades de los Gatos/microbiología , Gatos , Enfermedades de los Perros/microbiología , Perros , Infecciones por Bacterias Gramnegativas/epidemiología , Infecciones por Bacterias Gramnegativas/microbiología , Hungría/epidemiología , Incidencia , Neisseriaceae/aislamiento & purificación , Pasteurellaceae/aislamiento & purificación , Infecciones por Pasteurellaceae/epidemiología , Infecciones por Pasteurellaceae/microbiología , Infecciones por Pasteurellaceae/veterinaria , Faringe/microbiología , Sistema Respiratorio/microbiología , Enfermedades Respiratorias/epidemiología , Enfermedades Respiratorias/microbiologíaRESUMEN
In this paper we report the phenotypic and partial genetic characterisation of a novel bacterium strain isolated from a cat with severe nephritis. Multilocus sequence analysis was performed on the 16S rRNA and three housekeeping (recN, rpoB, infB) gene sequences obtained by PCR. In accordance with the results of phenotypic tests, the phylogenetic analyses confirmed the relatedness of the new strain (6036) to the family Pasteurellaceae. On the phylogenetic trees, strain 6036 appeared in a separate branch, closest to that of the type species (Frederiksenia canicola) of the genus Frederiksenia. These two bacteria shared 95.14 and 76.88% identity in their partial 16S rRNA and recN gene sequences, respectively. The rpoB- and infB-based phylogenetic analyses indicated that strain 6036 is most closely related to Bibersteinia trehalosi (with 90.58% identity) and [Haemophilus] felis ATCC 49733 (89.50% identity), respectively. The predicted genome identity values, based on the recN gene sequences, suggested that strain 6036 can be classified into the genus Frederiksenia as a novel species. A PCR method, specific to strain 6036, was developed to allow its rapid and accurate identification and differentiation from F. canicola and other species of Pasteurellaceae. The minimal inhibitory concentrations of 18 antimicrobial agents for strain 6036 were also determined.
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Enfermedades de los Gatos/microbiología , Nefritis/veterinaria , Pasteurellaceae/aislamiento & purificación , Animales , Gatos , Genes Bacterianos , Pruebas de Sensibilidad Microbiana/veterinaria , Nefritis/microbiología , Pasteurellaceae/clasificación , Filogenia , ARN Bacteriano/análisis , ARN Ribosómico 16S/análisisRESUMEN
In the recent years, African swine fever has become the biggest animal health threat to the swine industry. To facilitate quick genetic analysis of its causative agent, the African swine fever virus (ASFV), we developed a simple and efficient method for next generation sequencing of the viral DNA. Execution of the protocol does not demand complicated virus purification steps, enrichment of the virus by ultracentrifugation or of the viral DNA by ASFV-specific PCRs, and minimizes the use of Sanger sequencing. Efficient DNA-se treatment, monitoring of sample preparation by qPCR, and whole genome amplification are the key elements of the method. Through detailed description of sequencing of the first Hungarian ASFV isolate (ASFV_HU_2018), we specify the sensitive steps and supply key reference numbers to assist reproducibility and to facilitate the successful use of the method for other ASFV researchers.
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Virus de la Fiebre Porcina Africana/genética , Fiebre Porcina Africana/virología , Genoma Viral , Genómica , Secuenciación Completa del Genoma , Animales , ADN Viral , Genómica/métodos , Técnicas de Amplificación de Ácido Nucleico , Filogenia , PorcinosRESUMEN
Riemerella anatipestifer (RA) is a widely distributed bacterial pathogen of birds responsible for remarkable losses to poultry production, especially among waterfowl. We characterized the genomic diversity of 166 field isolates of RA, collected from geese and ducks, using enterobacterial repetitive intergenic consensus (ERIC)-polymerase chain reaction (PCR). The field strains and five reference strains showed 17 distinct patterns consisting of five to 12 bands ranging from approximately 150-1800bp. The majority of the strains belonged to two closely related ERIC-PCR types (A and B), while the other types contained only a few isolates each. There was no association between ERIC-PCR type and host species, place, or year of isolation; however the ERIC-PCR pattern was correlated with serotype for most isolates. The majority of serotype 1 strains (101/107) belonged to ERIC-PCR type A while the remaining six strains represented five different ERIC-PCR types (D, G, L, M, and O). Serotypes 1,7 and 7 corresponded to ERIC-PCR types B and C, respectively. Serotypes 2, 4, and 10 could be subdivided by ERIC-PCR revealing two to four patterns within each serotype. These results indicate that ERIC-PCR may be a suitable technique for the molecular identification of RA serotypes, and the detection of subtypes within certain serotypes may aid further epidemiological investigations. RESEARCH HIGHLIGHTS ERIC-PCR analysis of field R. anatipestifer strains revealed 17 distinct patterns Most strains belonged to two closely related ERIC-PCR types Serotype 1 was the most prevalent serotype representing 64.5% of the strains ERIC-PCR may be suitable for molecular identification of R. anatipestifer serotypes.
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Patos/microbiología , Infecciones por Flavobacteriaceae/veterinaria , Gansos/microbiología , Enfermedades de las Aves de Corral/microbiología , Riemerella/genética , Animales , Técnicas de Tipificación Bacteriana/veterinaria , Enterobacteriaceae/genética , Infecciones por Flavobacteriaceae/microbiología , Genotipo , Reacción en Cadena de la Polimerasa/veterinaria , Riemerella/inmunología , Riemerella/aislamiento & purificación , SerogrupoRESUMEN
Anatipestifer disease is a contagious disease caused by Riemerella anatipestifer, affecting primarily ducks, geese and turkeys, and characterised by listlessness, diarrhoea, sneezing, nasal discharge, and nervous signs. Sporadically, it occurs in a wide range of other domesticated and wild birds as well. The incidence and characteristics of the disease seen in the three main host species are summarised based on birds submitted for routine laboratory investigation in Hungary over the period 2010-2014. The infection was diagnosed in a higher percentage in geese (9.9%) and ducks (7.5%). It occurred in 5-day-old to 17-week-old geese and 3- to 6.5-week-old ducks, respectively. The pathological lesions were comparable in these two species: enlarged spleen, serofibrinous pericarditis, perihepatitis, airsacculitis, catarrhal enteritis, subcutaneous oedema and hyperaemia over the cranium, mucopurulent exudate in the nasal cavity and occasionally pneumonia, conjunctivitis, purulent arthritis and caseous salpingitis. In some cases, R. anatipestifer produced only secondary lesions, which complicated other diseases such as circovirus infection, mycotoxicosis, mycoplasmosis, or Derzsy's disease. In turkeys, the disease occurred rarely (0.5%) and at an older age (12 to 19 weeks). The lesions most frequently seen were purulent osteomyelitis of the cranium and seropurulent meningitis. Purulent osteomyelitis in the cranium caused by R. anatipestifer infection had not been reported in turkeys previously. To various extents, other local lesions such as serofibrinous pericarditis, airsacculitis, arthritis, and in one case septicaemia were also observed. The high incidence of the disease in waterfowl underlines the importance of appropriate treatment and prevention that should be based on accurate diagnosis and antimicrobial susceptibility testing, proper biosecurity and vaccination with regard to the serotype(s) present on the farm.
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Infecciones por Flavobacteriaceae/veterinaria , Enfermedades de las Aves de Corral/microbiología , Aves de Corral , Riemerella/clasificación , Animales , Infecciones por Flavobacteriaceae/epidemiología , Infecciones por Flavobacteriaceae/microbiología , Hungría/epidemiología , Enfermedades de las Aves de Corral/epidemiología , Factores de TiempoRESUMEN
The emergence of simultaneous resistance to multiple classes of antibiotics presents an increasing threat. Plasmid-borne multiresistance and integrative conjugative elements have been reported in Pasteurella multocida. We report an alternative strategy for the development of multiresistance observed in a P. multocida strain (Pm238) isolated from calf pneumonia. We identified genes integrated into the chromosomal DNA without known integrative and conjugative elements. These genes conferred resistance to streptomycin (strA), tetracycline (tetB), chloramphenicol (catAIII), and sulphonamides (sulII). We also detected mutation in the quinolone-resistance-determining regions of parC. No plasmids could be isolated from strain Pm238. These results suggest that P. multocida can accumulate multiple resistance determinants on the chromosome as single genes.
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Antibacterianos/farmacología , Enfermedades de los Bovinos/microbiología , Farmacorresistencia Bacteriana Múltiple , Infecciones por Pasteurella/veterinaria , Pasteurella multocida/efectos de los fármacos , Animales , Bovinos , Infecciones por Pasteurella/microbiologíaRESUMEN
Riemerella anatipestifer causes anatipestifer disease in many avian species. A total of 185 R. anatipestifer strains isolated in Hungary between 2000 and 2014 from geese and ducks were tested against 13 antibiotics (ampicillin, doxycycline, enrofloxacin, erythromycin, florfenicol, flumequine, gentamicin, penicillin, spectinomycin, streptomycin, sulphamethoxazole-trimethoprim, sulphonamide compounds, and tetracycline) by the Kirby-Bauer disk diffusion method. The majority of the strains were susceptible to florfenicol (97.9%), ampicillin (95.1%), penicillin (93%), sulphamethoxazole-trimethoprim (92.4%), and spectinomycin (86.5%). The highest resistance rates were observed for flumequine, tetracycline, erythromycin and streptomycin (94%, 91.4%, 75.1% and 71.4% resistance, respectively). The resistance patterns showed some variation depending on the geographical origin of the strains. The average rate of extensive drug resistance was 30.3%, and its proportion tended to increase in the period examined.
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Anseriformes , Antibacterianos/farmacología , Farmacorresistencia Bacteriana , Enfermedades de las Aves de Corral/microbiología , Riemerella/clasificación , Riemerella/efectos de los fármacos , Animales , Hungría/epidemiología , Enfermedades de las Aves de Corral/epidemiología , Riemerella/aislamiento & purificaciónRESUMEN
Ornithobacterium rhinotracheale is a widely distributed rod-shaped Gram-negative bacterium that infects several avian species including chickens and turkeys. It is associated with respiratory signs, growth retardation, mortality, and reduced egg production, thus causing severe economic losses to the poultry industries. In this study, 37 field isolates of O. rhinotracheale, collected from various locations in Hungary between 1997 and 2015, were identified and characterized by the analysis of partial 16S rRNA gene sequences, enterobacterial repetitive intergenic consensus (ERIC)-PCR, and random amplified polymorphic DNA (RAPD) PCR assays with the OPG11, OPH19, and M13 primers. Most of the field isolates were serotype A, one was serotype B, and four were serotype D. One isolate could not be typed with antisera against serotypes A-E. In a phylogenetic analysis of the 16S rRNA sequences, the isolates formed two clusters. Thirteen distinct patterns were identified with ERIC-PCR, and the RAPD assay with the M13 primer assigned the isolates to 10 different patterns. The other two RAPD assays were unsuitable for distinguishing and grouping the isolates. Neither ERIC type nor RAPD pattern correlated with the place or year of isolation. However, the strains isolated from chickens were more heterogeneous on ERIC-PCR than the isolates recovered from turkeys. In this study, ERIC-PCR was the most discriminatory method for investigating the genetic diversity of O. rhinotracheale isolates.