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      Caracterización microbiológica y genotípica de cepas de Pasteurella multocida asociadas al síndrome respiratorio cunícola Translated title: Microbiological and genotypic characterization of Pasteurella mutocida strains associated with the cunicular respiratory syndrome

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          Abstract

          RESUMEN Pasteurella multocida es una bacteria Gram negativa que se asocia a síndromes respiratorios en diferentes especies de animales. Esta bacteria presenta una alta heterogeneidad intraespecie, donde es posible encontrar diferentes biovares, cinco serogrupos capsulares con diferencias en los factores de virulencia y la expresión de la patogenicidad, así como en los perfiles de susceptibilidad a los antibióticos y la capacidad para la formación de biopelículas. El objetivo del presente trabajo es la caracterización de cepas de P. multocida recobradas de conejos. Se realizó la identificación microbiológica, la genoserotipificación capsular y la detección de genes que participan en la expresión de factores de virulencia. Se determinó la susceptibilidad a varias familias de antibióticos y la capacidad de producir biopelículas in vitro. Se confirmó que las cepas pertenecen a la especie P. multocida subespecie multocida biotipo 1, al tipo capsular A. Las cepas fueron positivas a la detección de fragmentos de genes de los locis relacionados con la biogénesis de fimbrias, neurominidasa y proteínas de membrana externa, que constituyen factores de virulencia, específicamente para la colonización e interacción con el hospedero. Las cepas revelaron multirresistencia y solo dos mostraron moderada capacidad para la producción de biopelículas. La caracterización de estas cepas esclarece aspectos epidemiológicos relacionados con el serogrupo capsular, que son de utilidad para la obtención de candidatos vacunales y contribuir a la prevención o profilaxis de infecciones por esta bacteria, por lo que el presente trabajo constituye una contribución al control de esta infección en conejos.

          Translated abstract

          ABSTRACT Pasteurella multocida is a Gram-negative bacterium associated with respiratory syndromes in different species of animals such as pigs, birds, rabbits, cats, and dogs. This bacterium presents high intra-species heterogeneity, where it is possible to find different biovars, five capsular serogroups with differences in virulence factors and the expression of pathogenicity, as well as profiles of susceptibility to antibiotics and the capacity for the formation of biofilms. The objective of this work is to characterize of P. multocida strains isolated from rabbits. The microbiological identification, capsular genoserotyping and the detection of genes involved in the virulence factors were carried out. Susceptibility to several families of antibiotics and the ability to produce biofilms in vitro were determined. It was confirmed that the strains belong to the species P. multocida subspecies multocida biotype 1 capsular A serogroup. The strains were positive to the detection of loci gene fragments related to the biogenesis of fimbrias, neurominidase and outer membrane proteins, which constitute virulence factors, specifically for colonization and interaction with the host. The strains revealed multi-resistance and only two of them showed moderate capacity for the production of biofilms. The characterization of these strains clarifies the epidemiological aspects related to the capsular serogroup, which are useful for obtaining vaccine candidates and contribute to the prevention or prophylaxis of infections by this bacterium, thus the present work contributes to the control of this infection in rabbits.

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          Antimicrobial Resistance in Bacterial Poultry Pathogens: A Review

          Antimicrobial resistance (AMR) is a global health threat, and antimicrobial usage and AMR in animal production is one of its contributing sources. Poultry is one of the most widespread types of meat consumed worldwide. Poultry flocks are often raised under intensive conditions using large amounts of antimicrobials to prevent and to treat disease, as well as for growth promotion. Antimicrobial resistant poultry pathogens may result in treatment failure, leading to economic losses, but also be a source of resistant bacteria/genes (including zoonotic bacteria) that may represent a risk to human health. Here we reviewed data on AMR in 12 poultry pathogens, including avian pathogenic Escherichia coli (APEC), Salmonella Pullorum/Gallinarum, Pasteurella multocida, Avibacterium paragallinarum, Gallibacterium anatis, Ornitobacterium rhinotracheale (ORT), Bordetella avium, Clostridium perfringens, Mycoplasma spp., Erysipelothrix rhusiopathiae, and Riemerella anatipestifer. A number of studies have demonstrated increases in resistance over time for S. Pullorum/Gallinarum, M. gallisepticum, and G. anatis. Among Enterobacteriaceae, APEC isolates displayed considerably higher levels of AMR compared with S. Pullorum/Gallinarum, with prevalence of resistance over >80% for ampicillin, amoxicillin, tetracycline across studies. Among the Gram-negative, non-Enterobacteriaceae pathogens, ORT had the highest levels of phenotypic resistance with median levels of AMR against co-trimoxazole, enrofloxacin, gentamicin, amoxicillin, and ceftiofur all exceeding 50%. In contrast, levels of resistance among P. multocida isolates were less than 20% for all antimicrobials. The study highlights considerable disparities in methodologies, as well as in criteria for phenotypic antimicrobial susceptibility testing and result interpretation. It is necessary to increase efforts to harmonize testing practices, and to promote free access to data on AMR in order to improve treatment guidelines as well as to monitor the evolution of AMR in poultry bacterial pathogens.
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            Isolation, antimicrobial resistance, and virulence genes of Pasteurella multocida strains from swine in China.

            A total of 233 isolates of Pasteurella multocida were obtained from 2,912 cases of clinical respiratory disease in pigs in China, giving an isolation rate of 8.0%. Serogroup A P. multocida isolates were isolated from 92 cases (39.5%), and serogroup D isolates were isolated from 128 cases (54.9%); 12 isolates (5.2%) were untypeable. P. multocida was the fourth most frequent pathogenic bacterium recovered from the respiratory tract, after Streptococcus suis, Haemophilus parasuis, and Escherichia coli. All isolates were characterized for their susceptibilities to 20 antibiotics and the presence of 19 genes for virulence factors (VFs). The frequency of antimicrobial resistance among P. multocida isolates from swine in China was higher than that reported among P. multocida isolates from swine in from other countries, and 93.1% of the isolates showed multiple-drug resistance. There was a progressive increase in the rate of multiresistance to more than seven antibiotics, from 16.2% in 2003 to 62.8% in 2007. The resistance profiles suggested that cephalosporins, florfenicol, and fluoroquinolones were the drugs most likely to be active against P. multocida. Use of PCR showed that colonization factors (ptfA, fimA, and hsf-2), iron acquisition factors, sialidases (nanH), and outer membrane proteins occurred in most porcine strains. The VFs pfhA, tadD, toxA, and pmHAS were each present in <50% of strains. The various VFs exhibited distinctive associations with serogroups: concentrated in serogroup A, concentrated in serogroup D, or occurring jointly in serogroups A and D. These findings provide novel insights into the epidemiological characteristics of porcine P. multocida isolates and suggest that the potential threat of such multiresistant bacteria in food-producing animals should not be neglected.
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              Virulence genotype of Pasteurella multocida strains isolated from different hosts with various disease status.

              To learn more about the molecular biology of Pasteurella multocida 289 strains isolated from various clinically healthy and diseased hosts were examined for capsule biosynthesis genes (capA, B, D, E, and F) and 14 virulence associated genes by PCR and DNA-DNA-hybridization. As expected, capsule type A strains were highly adapted to bovines (92.3%) and poultry (85.7%) while we mainly found capA (34.9%)- and capD (58.1%)-positive strains in swine. A noticeable amount of capD-positive strains also originated from small ruminants (34.9%) and capF was detected in wild type strains from diseased cattle (2.2%) and cats (7.4%). None of the isolates harboured capE, while capB was exclusively found in all strains from buffaloes. Nearly all isolates showed a combination of genes encoding outer membrane proteins, colonization factors, iron aquisition factors and superoxid-dismutases without any clue for host specificity. In contrast, the transferrin binding protein encoding gene tbpA (31.5%) was limited to ruminant strains and only 37.0% of all P. multocida strains harboured pfhA, coding for a filamentous hemagglutinin, supposed to be a putative adhesion- und serum resistance factor. PfhA revealed a strong positive association to the outcome of disease in bovine hosts and in combination with toxA to that in swine. The dermonecrotoxin encoding toxA, present in 12.5% of all strains, was detected in isolates from swine, small ruminants, cattle, and poultry. A significant association to the disease status, however, was only existent in swine, although with 66.7% we found a notably high prevalence of the toxin gene among strains from small ruminants. The genes toxA, tbpA and pfhA as well as capsule biosynthesis genes are supposed to be important epidemiological marker genes for characterizing P. multocida field strains.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                rsa
                Revista de Salud Animal
                Rev Salud Anim.
                Centro Nacional de Sanidad Agropecuaria (La Habana, , Cuba )
                0253-570X
                2224-4700
                April 2019
                : e02
                Affiliations
                [1] Boyeros La Habana orgnameCentro Nacional para la Producción de Animales de Laboratorio Cuba
                [2] San José de las Lajas La Habana orgnameCentro Nacional de Sanidad Agropecuaria Cuba
                Article
                S0253-570X2019000100003
                df76892f-6d8d-43cc-b14b-7c2614871541

                This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

                History
                : 16 November 2018
                : 21 January 2019
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 46, Pages: 0
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                SciELO Cuba


                biopelículas,factores de virulencia,Pasteurella multocida,resistencia antimicrobiana,serogrupo capsular,biofilms,virulence factors,antimicrobial resistance,capsular serogroup

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