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D'Silva, Colin Gerard. "Iron acquisition by Actinobacillus pleuropneumoniae." Thesis, McGill University, 1995. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=28720.
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Four strains of the swine pathogen Actinobacillus pleuropneumoniae, namely, the type strain (ATCC 27088), the "reference" strain of biotype 2 (Bertschinger 2008/76) and two additional biotype 1 strains, strain BC181, which is less virulent than the type strain, and strain K17 (reference strain of serotype 5A), which was isolated from a lamb, were investigated with respect to iron acquisition. All four strains produced iron-repressible outer membrane proteins. However, only strains ATCC 27088 and Bertschinger 2008/76 could acquire iron from porcine transferrin. No organism could utilize human, bovine or ovine transferrin, or ovine or porcine lactoferrin. Haemoglobin supported good growth of all strains except K17 (which also failed to acquire iron from haemin). In all cases, iron acquisition from transferrin or haemoglobin required direct contact between the organisms and the proteins. Total membranes derived from iron-restricted organisms were subjected to an affinity isolation technique based on biotinylated porcine transferrin and streptavidin-agarose, and the following polypeptides were isolated: 99 kDa and 64 kDa from strain ATCC 27088; 93 kDa from strain Bertschinger 2008/76; 95 kDa (trace amounts) and 60 kDa from strain BC181; none from strain K17. These polypeptides appear to be transferrin receptor components. The 99 kDa polypeptide (TBPl) from the type strain was purified by SDS-PAGE and transferred electrophoretically onto polyvinylidene difluoride membrane. The N-terminal amino acid sequence of the polypeptide was determined commercially. A commercially-synthesized oligonucleotide probe was used to clone the gene encoding the TBPl of the type strain in competent Escherichia coli DH5$ alpha$ cells.
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Maas, Alexander. "DIVA vaccine development against Actinobacillus pleuropneumoniae infection." [S.l.] : [s.n.], 2006. http://deposit.ddb.de/cgi-bin/dokserv?idn=983732574.
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MacKay, David Keith John. "Immunity to 'Actinobacillus (Haemophilus) pleuropneumoniae' in piglets." Thesis, Royal Veterinary College (University of London), 1993. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.519530.
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Ali, Tehmeena. "Characterisation of protein secretion systems of Actinobacillus pleuropneumoniae." Thesis, University of Nottingham, 2007. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.440122.
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Stäger, Martin. "Zur Seroprävalenz Actinobacillus pleuropneumoniae (APP) in Schweizer Schweinezuchtbeständen /." Bern, 1991. http://www.ub.unibe.ch/content/bibliotheken_sammlungen/sondersammlungen/dissen_bestellformular/index_ger.html.
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Silva, Thyara Ferreira da. "Caracterização e expressão da chaperona Hfq em Actinobacillus pleuropneumoniae, o agente causal da pleuropneumonia suína." Universidade Federal de Viçosa, 2016. http://www.locus.ufv.br/handle/123456789/10060.
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Conselho Nacional de Desenvolvimento Científico e Tecnológico
Infecções que acometem o trato respiratório de suínos levam a significativas perdas econômicas na suinocultura mundial. Um dos principais patógenos respiratórios em suínos é a bactéria Actinobacillus pleuropneumoniae, o principal agente causal da pleuropneumonia. Atualmente podem ser encontrados 16 sorotipos de A. pleuropneumoniae com virulência distinta e complexa, sendo os principais fatores de virulência: exotoxinas Apx, lipopolissacarídeo (LPS), cápsula e a capacidade de formação de biofilme. O controle da doença é baseado no uso de antibióticos e cuidados no manejo da granja. A profilaxia pela imunização passiva ainda é ineficiente devido à dificuldade na obtenção de uma vacina contra todos os sorotipos encontrados. Assim, novas abordagens experimentais na elaboração de uma vacina eficiente associada a uma resposta imune protetora são essenciais porque podem representar novas alternativas na estratégia de controle da doença. A proteína Hfq é um componente central de regulação global pós-transcricional e participa diretamente na regulação da expressão de genes por facilitar a interação de RNAs pequenos com mRNAs alvos, sendo esta uma abordagem atual e relacionada ao controle da virulência em diversas bactérias patogênicas. Neste sentido, o estudo da chaperona de RNA Hfq é de extrema importância, uma vez que já foi demonstrado seu efeito pleiotrópico e impacto na virulência, na resposta a diferentes tipos de estresse e no crescimento celular de vários patógenos, incluindo A. pleuropneumoniae. Portanto, esse trabalho teve como objetivos: caracterizar in silico a proteína Hfq em A. pleuropneumoniae e analisar a expressão e a fase de maior abundância desta proteína ao longo do crescimento de A. pleuropneumoniae. As análises filogenéticas realizadas foram baseadas em análises de comparação de sequências de aminoácidos da proteína Hfq de diferentes membros da classe Gammaproteobacteria, na qual A. pleuropneumoniae está inserida. As demais análises foram conduzidas utilizando os sorotipos 1, 8 e 15 de A. pleuropneumoniae, sendo utilizadas as linhagens do tipo selvagem (WT) e hfq::3XFLAG. O alinhamento de sequências da proteína Hfq revelou uma identidade de 98% entre as proteínas Hfq de A. pleuropneumoniae de diferentes sorotipos, além de demonstar que Hfq de espécies de uma mesma família possuem maior relação filogenética. A análise da estrutura tridimensional da proteína em A. pleuropneumoniae demonstrou a presença de estruturas características da proteína presente em outos patógenos Gram-negativos, como uma α-hélice e folhas β. A análise da velocidade específica de crescimento por ANOVA entre as linhagens WT e hfq::3XFLAG do mesmo sorotipo revelou que não há diferença de crescimento entre essas linhagens do mesmo sorotipo. Quanto à expressão de Hfq, foi detectado um maior acúmulo da proteína na fase estacionária de crescimento, no período de 6-8 horas, dependendo do sorotipo investigado, e que a expressão de Hfq foi diferencial entre os sorotipos analisados. Esses resultados revelaram que a proteína Hfq é conservada entre os sorotipos de A. pleuropneumoniae e possui estrutura tridimensional característica. Além disso, a inserção da etiqueta FLAG em Hfq não alterou o perfil de crescimento celular e hà um maior acúmulo da proteína na fase estacionária de crescimento, sendo que os sorotipos apresentaram distribuição das formas diferencial entre os sorotipos e dinâmica de acordo com a fase de crescimento. Essa diferença pode estar relacionada aos diferentes perfis de virulência e de resposta a diferentes condições investigadas previamente, uma vez que a abundância nestes sorotipos apresentou distribuição temporal distinta.
Infections that affect the respiratory tract of pigs lead to significant economic losses in the swine industry worldwide. One of the major respiratory pathogen in pigs is the bacterium Actinobacillus pleuropneumoniae, the main causal agent of the pleuropneumonia. Currently, 16 serotypes can be found of A. pleuropneumoniae with distinct and complex virulence, with the main factors of virulence: exotoxin Apx, lipopolysaccharide (LPS), capsule and the biofilm formation capacity. Control of the disease is based on the use of antibiotics and care in the management of the farm. Prophylaxis by passive immunization is still inefficient because of the difficulty in getting a vaccine against all serotypes found. Thus, new experimental approaches in the development of an effective vaccine associated with a protective immune response are essential because they can represent new alternatives in the disease control strategy. The Hfq protein is a key component of the global post-transcriptional regulation and directly participates in the regulation of gene expression to facilitate the interaction of small RNAs with target mRNAs, which is a current approach and it relates to the control of virulence in many pathogenic bacteria. In this sense, the study of Hfq RNA chaperone is of extreme importance, since it has already demonstrated its pleiotropic effect and impact on virulence in response to different types of stress and cellular growth of various pathogens, including A. pleuropneumoniae. Therefore, this study aimed: to characterize in silico the Hfq protein in A. pleuropneumoniae and to analyze the expression and phase greater abundance of this protein throughout the growth of A. pleuropneumoniae. The phylogenetic analyzes were based on comparative analysis of amino acid sequences of protein Hfq of different members of the class Gammaproteobacteria, which A. pleuropneumoniae is inserted. The other analyzes were conducted using the serotypes 1, 8 and 15 of A. pleuropneumoniae, being used strains of wild-type (WT) and hfq::3XFLAG. The sequence alignment of Hfq protein sequences showed an identity of 98% between Hfq proteins of A. pleuropneumoniae of different serotypes, also demonstrating that Hfq species of the same family have a greater phylogenetic relationship. The analysis of the three-dimensional structure of the protein in A. pleuropneumoniae demonstrated the presence of specific structures of protein present in other Gram-negative pathogens, how one α-helix and β-strands. The analysis of the specific growth rate by ANOVA between strains WT and hfq::3XFLAG of the same serotype showed that there is no difference in growth between the strains. As the expression of Hfq, a greater accumulation of protein in the stationary growth phase was detected in the period of 6-8 hours, depending on the serotype investigated, and the expression of Hfq was differential between serotypes analyzed. These results demonstrate that Hfq is conserved among serotypes of A. pleuropneumoniae and has a three-dimensional structure conserved. Moreover, the insertion of the FLAG tag on Hfq did not affect the cell growth profile and there is a greater accumulation of the protein in the stationary phase of growth, whereas serotypes showed the distribution of forms differential between serotypes and dynamically according to the growth stage. This difference may be related to different profiles of virulence and response to different conditions previously investigated, since these abundant serotypes showed distinct temporal distribution.
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Ma, Jianneng. "Purification, serology and pathogenic role of the 110 kilodalton rtx hemolysins of Actinobacillus pleuropneumoniae." Diss., Virginia Tech, 1991. http://hdl.handle.net/10919/39935.
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Actinobacillus pleuropneumoniae is the etiological agent of contagious swine pleuropneumonia, an economically important disease of the swine industry worldwide. Improved control of this disease requires enhanced understanding of the factors contributing to pathogenesis. The objectives of this study were to investigate the immune response and virulence properties of the 110-kilodalton (110-KDa) hemolysins [hemolysin I (HlyI) and hemolysin II (HlyII)] of A. pleuropneumoniae. Several monoclonal antibodies (MAb) to the hemolysins were developed. An IgGl. MAb (8C2) specific for HlyII, as determined by immunoblotting, was cross-linked to Protein A-Sepharose, and HlyII was purified from serotypes 1 and 5 by immunoaffinity chromatography. An indirect enzyme-linked immunosorbent assay (ELISA) using MAb 8C2, or affinitypurified rabbit IgG to both hemolysins, was developed for detection of swine antibody to one or both hemolysins, respectively. In comparison with the complement fixation test, the ELISA was highly sensitive and specific, and was able to identify animals infected with or exposed to most, if not all, serotypes of A. pleuropneumoniae. Several nonhemolytic mutants of A. pleuropneumoniae serotype 5 were isolated following electroporation of the parent with an hemolysin gene whose open-reading-frame was disrupted with a kanamycin resistance gene. One mutant was characterized for phenotypic and pathogenic properties. Biochemical profiles, growth rate, capsule content, and lipopolysaccharide and whole cell protein electrophoretic profiles of the parent and one of the mutants were similar. The nonhemolytic mutant lacked both HlyI and HlyII proteins in culture supernatant and in whole cell lysates as determined by immunoblot analysis; extracellular and intracellular hemolytic and cytotoxic activity was also absent. The mutant was avirulent in mice and pigs at doses greater than 10 times the lethal dose of the parent. Unlike the parent, the nonhemolytic mutant failed to confer protection against lethal challenge in mice following immunization. Thus, one or both hemolysins are essential for virulence and immunoprotection in A. pleuropneumoniae serotype 5.Ph. D.
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Carufel, Karine de. "Mutagenèse par la technologie du transposome chez Actinobacillus pleuropneumoniae /." Thèse, Trois-Rivières : Université du Québec à Trois-Rivières, 2008. http://www.uqtr.ca/biblio/notice/resume/30037972R.pdf.
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Carufel, Karine de. "Mutagenèse par la technologie du transposome chez Actinobacillus pleuropneumoniae." Thèse, Université du Québec à Trois-Rivières, 2008. http://depot-e.uqtr.ca/1839/1/030037972.pdf.
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Costa, Bárbara Letícia Pereira. "Caracterização fenotípica e genotípica de isolados de Actinobacillus pleuropneumoniae provenientes de diferentes estados brasileiros." Universidade de São Paulo, 2017. http://www.teses.usp.br/teses/disponiveis/10/10134/tde-27072017-151034/.
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A infecção por Actinobacillus pleuropneumoniae, doença conhecida como pleuropneumonia suína, assumiu grande importância na suinocultura moderna devido à alta ocorrência observada nos rebanhos. O impacto da doença está relacionado à capacidade do agente em causar pneumonia severa, levando os animais a óbito ou doença crônica, resultando em graves prejuízos zootécnicos. Diante desse cenário, o controle e monitoramento do agente se faz importante por meio da identificação dos diferentes sorotipos, da análise genética e da determinação dos perfis de resistência aos antimicrobianos. O objetivo do presente estudo foi caracterizar fenotípica e genotípicamente estirpes de Actinobacillus pleuropneumoniae isoladas a partir de quadros de pneumonia em suínos. Um total de 85 estipes de A. pleuropneumoniae foram submetidas a reação em cadeia pela polimerase (PCR) para identificação e sorotipagem, determinação da concentração inibitória mínima de antimicrobianos, polimorfismo do comprimento de fragmentos amplificados (AFLP) e eletroforese em gel de campo pulsado (PFGE). Os sorotipos mais frequentes foram: 5 (38,8%), 10 (29,4%), 7 (5,9%), 8 (5,9%) e 6 (3,5%), sendo que 14 (16,5%) estirpes foram não tipáveis. Foi observada alta heterogeneidade de perfil genético entre as estirpes analisadas, tanto pelo AFLP quanto pelo PFGE, e o índice discriminatório para cada técnica foi 0,97 e 0,84, respectivamente. Todas as estirpes foram sensíveis ao ceftiofur, gentamicina, tulatromicina e tilmicosina, sendo que 98,8% das estirpes foram resistentes à tilosina e altas taxas de resistência foram observadas ainda para as tetraciclinas, clindamicina e sulfadimetoxina.Infection by Actinobacillus pleuropneumoniae, a disease known as swine pleuropneumonia, has gained greater relevance to modern pig farming due to the high recurrence rate observed in herds. The impact of the disease relates to the capacity of the agent to cause severe pneumonia, leading to animal death or chronic conditions, thus resulting in severe zootechnical losses. In view thereof, the control and monitoring of the agent is key, being performed through the identification of different serotypes, genetic analysis and determination of antimicrobial resistance profiles. The objective of this study was to characterize phenotypically and genotypically Actinobacillus pleuropneumoniae strains isolated from swine with clinical presentation of pneumonia. A total of 85 strains of A. pleuropneumoniae were subject to polymerase chain reaction (PCR) for identification and serotyping, determination of the minimal inhibitory concentration, amplified fragment length polymorphism (AFLP) and pulsed field gel electrophoresis typing techniques (PFGE). Most recurring serotypes were: 5 (38.8%), 10 (29.4%), 7 (5.9%), 8 (5.9%) and 6 (3.5%), of which 14 (16.5%) strains were nontypeable. High genetic heterogeneity was observed for both AFLP and PFGE, and the discriminatory index for each technique was 0.97 and 0.84, respectively. All 85 strains were susceptible to ceftiofur, gentamicin, tulatromicin and tilmicosin, 84 of which were resistant to tylosin, and high resistance rates were also observed for clindamycin, tetracyclines and sulfadimethoxine.
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Archambault, Marie. "Étude de la liaison de l'hémoglobine porcine chez Actinobacillus pleuropneumoniae." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 2000. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape2/PQDD_0026/NQ52124.pdf.
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Ricard, Michelle. "Iron acquisition from porcine proteins by Actinobacillus pleuropneumoniae biotype 1." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1999. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape2/PQDD_0034/MQ64438.pdf.
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Donovan, Elizabeth Anne. "Identification and characterisation of immunogenic vaccine candidates of 'Actinobacillus pleuropneumoniae'." Thesis, University of Nottingham, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.435764.
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Nahar, Nusrat. "Characterisation of gene expression and virulence factors in Actinobacillus pleuropneumoniae." Thesis, Griffith University, 2023. http://hdl.handle.net/10072/421340.
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Actinobacillus pleuropneumoniae is a Gram-negative bacterium causing the highly infectious disease porcine pleuropneumonia and is responsible for global financial losses to the swine industry every year. Though the virulence of A. pleuropneumoniae is complex and multifactorial, Apx toxins (ApxI-III) are the major contributing factors that causes lung lesions in pigs. Although vaccines are available to prevent A. pleuropneumoniae infections, they do not give complete protection and typically give protection against the serovars used to prepare the vaccine. Thus, a thorough understanding of gene expression and virulence factors is required to develop broadly protective pleuropneumonia vaccines.
This thesis first investigated a novel pathway to prevent and treat pleuropneumonia infection by blocking the interaction between Apx toxins and the host cells. To determine the specific ligands bound by each Apx toxin, glycan array analysis using purified Apx toxins (ApxI-III, both the active and inactive forms e.g. ApxCA and ApxA) was carried out. Expressing both with and without ApxC allowed an assessment of whether this activation is required for interaction with the host glycan receptor. Significant work was needed to optimise overexpression and purification of Apx toxins. Glycan array analysis demonstrated that both ApxI and ApxII toxins bound to very similar glycan structures, such as gangliosides and Lewis antigens. Binding of Apx toxins occurred irrespective of activation by the cognate acyltransferase, ApxC. Interestingly, glycan binding was not observed for the ApxIII toxin, indicating that interaction of this toxin with its already characterised host cell receptor, the CD18 subunit of β2 integrin, likely does not occur via glycan interactions. In recent years, systems known as phasevarions – for phase-variable regulons – have been described in multiple host-adapted bacterial pathogens. Phasevarions result from the rapid and reversible expression of genes encoding cytoplasmic DNA methyltransferases. This results in variable expression of these methyltransferases in a population, with variable genome wide methylation differences within a bacterial population resulting in differential expression of multiple genes via epigenetic mechanisms. In all described cases, phasevarions control expression of current and putative vaccine candidates. The study in Chapters 4 and 5 characterised the phase-variable Type I and Type III R-M systems identified in A.
pleuropneumoniae. A study of the distribution of both systems using 210 whole genome sequences demonstrated that the Type I R-M system is present in almost all serovars of A. pleuropneumoniae, whereas different phase-variable Type III mod genes showed colocalisation with specific serovars of A. pleuropneumoniae. This study also demonstrated that individual strains of A. pleuropneumoniae could encode both phase-variable Type I and Type III R-M systems, a phenomenon never before observed. In Chapter 4, phase-variable expression of the Type I R-M system was demonstrated using two prototype strains with a combination of semi-quantitative RT-PCR and a locus specific FAM-labelled PCR assay. The work carried out here also developed a method by which locked strains could be generated that only express a single HsdS variant.
Characterisation of the Type III mod genes in Chapter 5 revealed the presence of two distinct phase-variable Type III methyltransferases, modP (which exists in four variants, designted modP1 to modP4) and modQ in A. pleuropneumoniae. The serovar-specific distribution of each of these new mod genes was further confirmed using a second strain collection, comprising 265 strains from the Australian national culture collection. This study demonstrated that these Type III methyltransferases are phase-variable, and that each variant methylates a different DNA target sequence (established using a combination of Pacific Biosciences Single- Molecule, Real-Time (SMRT) sequencing and Oxford Nanopore sequencing). This methylome analysis demonstrated the presence of the first phase-variable cytosine-specific Type III DNA methyltransferase discovered in bacteria. Analysis of changes in gene expression and phenotype influenced by phase variation of each Type III methyltransferase showed that each distinct variant regulates different phasevarions, and has a unique influence on bacterial phenotype, such as antibiotic resistance (modP2), biofilm formation (modP1) and growth rate (modP1).
In summary, this thesis has taken the first step to both treating and preventing disease caused by A. pleuropneumoniae by characterising the exact host receptors bound by the major virulence factors ApxI and ApxII and will allow the development of new treatment strategies that can block the interaction between Apx toxins and their cellular receptors. This would allow for more effective treatments and negate the use of antibiotics for the treatment of porcine pleuropneumonia. The characterisation of the phase-variable Type I and Type III DNA methyltransferases described in this study has begun to define the stably expressed antigenic repertoire of A. pleuropneumoniae. This will direct and inform the development of rationally designed vaccines to prevent disease caused by this major veterinary pathogen.Thesis (PhD Doctorate)
Doctor of Philosophy (PhD)
Institute for Glycomics
Science, Environment, Engineering and Technology
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Brandão, Laila Natasha Santos. "Utilização da técnica de nanopartículas de ouro não modificada (AuNP) para detecção do agente da pleuropneumonia suína (Actinobaccilus pleuropneumoniae)." Universidade Federal de Mato Grosso, 2014. http://ri.ufmt.br/handle/1/534.
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CAPES
A possibilidade de detecção de agentes patogênicos representou um grande passo para a ciência. O desenvolvimento e melhorias nas técnicas de diagnóstico como a técnica de Reação em Cadeia da Polimerase (PCR), ao longo dos anos ainda requer infraestrutura laboratorial, apesar de sua sensibilidade e custo decrescente, investimentos em equipamentos e cuidados com fontes de contaminação externa. Técnicas que possam ser executas com facilidade e pouca mão de obra ou exigência de pessoas capacitadas, têm grande possibilidades de aplicabilidade. Neste estudo desenvolveu-se uma técnica de rápida execução e baixo custo, para detecção de um dos principais agentes causadores de pneumonias em granjas de suínos o Actinobaccilus pleuropneumoniae, com sensibilidade 93,8% e especificidade de 84,6% em amostras de pulmões com e sem lesão. O teste Kappa entre a PCR e nanopartícula de ouro foi 0,684 representando boa concordância. A técnica pode ser utilizada como alternativa aos testes convencionais, já que é de fácil e rápida execução e não exige infraestrututra e mão de obra especializada.
The possibility of detection of pathogens was a major step for science as a whole, developing and improving these techniques over the years to increase visibly. The development of the technique of Polymerase Chain Reaction ( PCR ) , although its sensitivity and decreasing cost over the years, it's still a handy little technique that requires laboratory infrastructure, high investments in equipment and care with sources of external contamination. Techniques that can be performed through the ease and little manpower or requirement of skilled people, always have great scope of applicability. This study develops a technique for quick and low- cost detection of a major causative agent of pneumonia in swine herds the Actinobaccilus pleuropneumoniae , with 93.8 % sensitivity and 84.6% specificity in samples of lungs with and without injury, the Kappa test between PCR and gold nanoparticle was 0,684 representing good agreement . The technique can be used as an alternative to conventional tests, since it is easy and quick to implement and does not require infrastructure and skilled labor.
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Schuchert, Jennifer Ann. "Detection of Actinobacillus Pleuropneumoniae and Identification of Serotypes 1, 2, and 8 by Multiplex Polymerase Chain Reaction." Thesis, Virginia Tech, 2002. http://hdl.handle.net/10919/34135.
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Traditional immunological assays used to serotype Actinobacillus pleuropneumoniae have been problematic due to cross- reactivity between serotypes, particularly serotypes 6 and 8. To avoid these serological cross-reactions, a multiplex PCR assay was developed to detect A. pleuropneumoniae and identify serotypes 1, 2, and 8. Primers specific to the conserved capsular polysaccharide export region of A. pleuropneumoniae serotype 5 amplified a 880 bp fragment in all serotypes excluding serotype 4 or a 489 bp DNA fragment in all serotypes including serotype 4. Primers specific to the capsular polysaccharide biosynthesis regions of A. pleuropneumoniae serotypes 1, 2, and 8 amplified a 1.6 kb, a 1.7 kb, and 970 bp fragment in the respective serotype. This PCR assay detects A. pleuropneumoniae and identifies serotypes 1, 2, and 8.Master of Science
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Lo, Terry. "Detection and Identification of Acinobacillus pleuropneumoniae serotype 5 by multiplex Polymerase Chain Reaction." Thesis, Virginia Tech, 1997. http://hdl.handle.net/10919/36902.
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Traditional serologic assays of Actinobacillus pleuropeumoniae often have
problems with cross-reactivity. To avoid the complications of antibody-antigen
reactions, a PCR assay was developed to detect Actinobacillus pleuropneumoniae
and identify serotype 5 strains. Primers specific to the conserved capsular export
region of A. pleuropneumoniae amplified a 0.7 kb DNA band in all strains with the
exception of serotype 4. A second set of primers specific to the unique capsular
biosynthesis region of serotype 5 amplified a unique 1.1 kb band for serotype 5
only. The sensitivity of this assay was determined to be less than 100 colony
forming units. This PCR assay enables us to detect A. pleuronpeumoniae and definitively
distinguishes serotype 5 strains from other serotyes.Master of Science
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Sunn, Kyaw. "Construction and characterization of genetically defined metabolic mutants of Actinobacillus pleuropneumoniae." [S.l.] : [s.n.], 2003. http://deposit.ddb.de/cgi-bin/dokserv?idn=970211775.
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Vaz, Clarissa Silveira Luiz. "Genotipificação de amostras sorotipificáveis e não sorotipificáveis de Actinobacillus pleuropneumoniae através de RAPD." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2002. http://hdl.handle.net/10183/2556.
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Actinobacillus pleuropneumoniae é o agente etiológico da pleuropneumonia suína, enfermidade amplamente distribuída no rebanho suíno mundial, responsável por prejuízos econômicos relevantes. Possui 12 sorotipos, determinados por técnicas de sorotipificação. Além disso é descrita a ocorrência de amostras não sorotipificáveis. O conhecimento do sorotipo prevalente nos surtos da enfermidade é necessário aos programas de profilaxia. Procurando contornar as dificuldades normalmente encontradas na sorotipificação de A. pleuropneumpnoae, a técnica de RAPD foi avaliada na genotipificação de amostras sorotipificáveis e não sorotipificáveis do agente. Foram utilizados amostras ATCC dos 12 sorotipos e amostras dos sorotipos, 1, 3, 5a, 5b, 7, 11 e 12 isolados no Brasil. Os primers OPG e OPG-19, utilizados individualmente nas reações, foram mais adequados para a diferenciação dos sorotipos. O primer OPG-19 detectou polimorfismos semelhantes entrer os sorotipos 1, 3, 4, 5 e 11; e sorotipos 7 e 12. O perfil de RAPD detectado pelo primier OPGF-10 diferenciou os isolados de campo dos sorotipos 1, 7, 11 e 12. Os sorotipos 3 e 5 apresentaram padrão de RAPD semelhantes, sendo diferenciados pelo perfil de exotoxinas característico, determinado previamente através de PCR. Este primer identificou quatro diferentes perfis de RAPD no sorotipo 3. Um destes foi semelhante ao obtido como sorotipo 11. Neste isolado, foi detecta a presença dos genes para ApxI e ApxII, características do sorotipo 11. As amostras do sorotipo 4 apresentaram perfil de RAPD semelhante ao identificado nos sorotipos 3 ou 5 com o primeir OPG-10, sendo identificada, por PCR, a presença dos genes para ApxI e ApxI, os quais não são característicos do sorotipo. Estas amostras foram isoladas em anos posteriores à amostras dos sorotipos 3 e 5 analisadas. Foi possível caracterizar 14 das 14 amostras não sorotipificáveis de A.pleuropneumpniae obtidas de suínos com sinais da doença. Entre as 4 amostras não sorotipificáveis isoladas de leitões sem sinais clínicos, apenas uma foi caracterizada através de RAPD. É possível que as demais amostras sejam outrtas bactérias NAD-dependente isoladas do trato respiratório de suínos. Amostras caracterizadas como A. minor e A. indolicus apresentaram perfis de RAPD divergentes dos identificados em isolados puros de A. pleuropneumoniae, comprovando a capacidade da técnica na caracterização do agente. Diferentes amostras do mesmo sorotipo de A. pleuropneumoniae apresentaram polimorfismos de RAPD idênticos, demonstrando reprodutividade da técnica. Os resultados comprovam a capacidade de tipificação de A. Pleuropneumoniae através de RAPD. A pesquisa de primers adequados para a diferenciação dos sorotipos 3, 4 e 5 aprimorar sua caracterização, o que pode vir a contribuir com as técnicas de sorotipificação tradicionalmente utilizados, ou permitir o uso como método de confirmação nas amostras cuja sorotipificação é problemática.
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Khan, Shamila. "Therapeutic effect of Interlenkin-4 and Interleukin-1 receptor antagonist in Actinobacillus pleuropneumoniae challenged pigs." University of Sydney. Anatomy and Pathology, 2005. http://hdl.handle.net/2123/625.
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Immunological stressors, in the form of clinical and sub-clinical disease are currently controlled using both prophylactic antibiotics in-feed, and therapeutic antibiotic treatment. Respiratory disease, primarily Actinobacillus pleuropneumoniae (App) infection, is recognised as a major factor causing reduced productivity in pigs. This thesis reports investigations into the use of novel immunomodulators in particular Interleukin 4 (IL-4) and Interleukin 1 receptor antagonist (IL-1ra) as alternatives to antibiotics to treat App infection. Immunological and molecular biological assays were used to investigate and accumulate data. An in vitro study undertaken to find potential anti-inflammatory substances, revealed that Interleukin 8 (IL-8) mRNA production stimulated by PMA or LPS in whole pigs' blood was suppressed by IL-4. IL-1ra also suppressed stimulated IL-8 mRNA production by heat killed App bacteria (KB) in vitro. An acute LPS challenge in pigs in vivo however, showed no variation in illness or weight loss between pigs treated prophylactically with anti-inflammatory substance (IL-4 and IL-1ra) and saline treated pigs. The use of plasmids as a delivery system for anti-inflammatory substance did not show promise since it did not enhance growth or prolong the expression of the substances in the pigs. However, in the chronic App challenge model IL-4 and IL-1ra administered prophylactically in vivo showed an ability to improve growth. The therapeutic administration of IL-4 and IL-1ra to App challenged pigs showed no difference in pigs' growth, regardless of the treatment or control administered. To conclude, IL-4 and IL-1ra showed promise when administered prophylactically and improved growth and abrogated disease under conditions of App challenge. However when IL-4 and IL-1ra where administered therapeutically they did not perform as well. Moreover these compounds have potential as a commercial application to reduce the growth reduction caused by disease such as App.
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Durham, Andrew Leonard. "Identification of the regulators of in vivo expressed genes in Actinobacillus pleuropneumoniae." Thesis, Imperial College London, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.428545.
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Mikael, Leonie G. "Identification and characterization of the ferric hydroxamate uptake receptor in actinobacillus pleuropneumoniae." [Montréal] : Université de Montréal, 2002. http://wwwlib.umi.com/cr/umontreal/fullcit?pNQ80465.
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Thèse (Ph. D.)--Université de Montréal, 2003."NQ-80465." "Thèse présentée à la faculté des études supérieures en vue de l'obtention du grade de philosophiae doctor (Ph. D.) en microbiologie et immunologie." Version électronique également disponible sur Internet.
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Pereira, Monalessa Fábia. "Uso de Galleria mellonella como modelo de infecção e estudo de fatores relacionados com a virulência de Actinobacillus pleuropneumoniae." Universidade Federal de Viçosa, 2015. http://www.locus.ufv.br/handle/123456789/8477.
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Conselho Nacional de Desenvolvimento Científico e Tecnológico
Actinobacillus pleuropneumoniae é o agente etiológico da pleuropneumonia suína, uma severa enfermidade que acomete suínos de todas as idades, gerando perdas econômicas significativas para a suinocultura mundial. Embora os 15 sorotipos conhecidos dessa bactéria possam causar a doença, existem diferenças marcantes de virulência entre eles. A virulência de A. pleuropneumoniae é multifatorial e está relacionada à composição e estrutura de polissacarídeos da cápsula, LPS, e toxinas da família RTX, além desses fatores, a aderência em forma de biofilme e a resistência a agentes antimicrobianos podem ser determinantes para virulência. Este trabalho estabeleceu um modelo de infecção alternativo para o estudo de A. pleuropneumoniae, utilizando larvas de Galleria mellonella e, posteriormente esse modelo foi usado para investigar a virulência de isolados clínicos de A. pleuropneumoniae sorotipo 8. Os mesmos isolados foram avaliados quanto ao potencial de formação de biofilme e resistência a antimicrobianos comumente empregados em campo. A partir dessas informações, isolados clínicos com diferenças significativas na virulência, no potencial de formação de biofilme e no perfil de resistência foram selecionados para o sequenciamento genômico. Os resultados mostraram que o modelo de infecção A. pleuropneumoniae – G. mellonella é capaz de diferenciar níveis de virulência de isolados clínicos de mesmo sorotipo, além de permitir a avaliação da eficiência de agentes antimicrobianos contra este patógeno. O modelo também mostrou eficiência para diferenciar virulência entre linhagens selvagem e mutante da mesma bactéria. Uma análise de correlação entre os dados de virulência, formação de biofilme e resistência a antimicrobianos permitiu que seis isolados fossem selecionados para o sequenciamento. Com a montagem e anotação foi possível verificar que os genomas de A. pleuropneumoniae sorotipo 8 apresentam tamanho de 2,2 ± 0,004 Mpb, com o conteúdo GC de 40,33% ± 0,263 e regiões codificadoras com uma média de tamanho de 817,3 ± 6,8 pb. As regiões codificadoras correspondem a 89,05% ± 0,13 do genoma, das quais a maior parte foi anotada como genes funcionais, o que permitirá a realização de estudos comparativos. Estes genomas apresentam em média 79,5 ± 24,05 genes exclusivos, revelando a alta variabilidade genética dessa espécie, que pode estar relacionada com a variação da virulência entre os isolados estudados.
Actinobacillus pleuropneumoniae is the etiological agent of porcine pleuropneumonia, a severe disease that affects pigs of all ages, causing significant economic losses to the swine industry worldwide. Although the 15 serotypes of this bacterium are known to cause the disease, there are marked differences in virulence between them. A. pleuropneumoniae virulence is multifactorial and involves capsular polysaccharides, LPS, and toxins of the RTX family. In addition to these factors, the adhesion in biofilm form and resistance to antimicrobial agents may be determinant for virulence. This work has established an alternative infection model for the study of A. pleuropneumoniae, using larvae of Galleria mellonella and this model was subsequently used to investigate the virulence of clinical isolates of A. pleuropneumoniae serotype 8. The same isolates were evaluated for biofilm formation potential and resistance to antimicrobials commonly used in the field. From this information, clinical isolates with significant differences in virulence, biofilm formation potential and resistance profile were selected for genomic sequencing. Results show that the A. pleuropneumoniae - G. mellonella infection model is capable of differentiating levels of virulence of clinical isolates of the same serotype. Furthermore, it can be used to evaluate the effectiveness of antimicrobial agents against this pathogen. The model also showed efficiency to differentiate the virulence between wild and mutant strains of the same bacteria. A correlation analysis between the virulence data, biofilm formation and antibiotic resistance allowed six isolates to be selected for genome sequencing. With the assembly and annotation, we found that the genomes of A. pleuropneumoniae serotype 8 present size of 2.2 ± 0.004 Mpb, with GC content of 40.33% ± 0.263 and coding regions with an average size of 817.3 ± 6.8 bp. The coding regions correspond to 89.05 ± 0.13% of the genome, most of which was recorded as functional genes, enabling the comparative studies with these genomes. These genomes have 79.5 ± 24.05 exclusive proteins, revealing the high genetic variability of the species, which may be related to the variation in virulence between these isolates.
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Khan, Shamila. "Therapeutic effect of Interleukin-4 and Interleukin-1 Receptor Antagonist in Actinobacillus pleuropneumoniae challenged pigs." Thesis, The University of Sydney, 2005. http://hdl.handle.net/2123/625.
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Immunological stressors, in the form of clinical and sub-clinical disease are currently controlled using both prophylactic antibiotics in-feed, and therapeutic antibiotic treatment. Respiratory disease, primarily Actinobacillus pleuropneumoniae (App) infection, is recognised as a major factor causing reduced productivity in pigs. This thesis reports investigations into the use of novel immunomodulators in particular Interleukin 4 (IL-4) and Interleukin 1 receptor antagonist (IL-1ra) as alternatives to antibiotics to treat App infection. Immunological and molecular biological assays were used to investigate and accumulate data. An in vitro study undertaken to find potential anti-inflammatory substances, revealed that Interleukin 8 (IL-8) mRNA production stimulated by PMA or LPS in whole pigs' blood was suppressed by IL-4. IL-1ra also suppressed stimulated IL-8 mRNA production by heat killed App bacteria (KB) in vitro. An acute LPS challenge in pigs in vivo however, showed no variation in illness or weight loss between pigs treated prophylactically with anti-inflammatory substance (IL-4 and IL-1ra) and saline treated pigs. The use of plasmids as a delivery system for anti-inflammatory substance did not show promise since it did not enhance growth or prolong the expression of the substances in the pigs. However, in the chronic App challenge model IL-4 and IL-1ra administered prophylactically in vivo showed an ability to improve growth. The therapeutic administration of IL-4 and IL-1ra to App challenged pigs showed no difference in pigs' growth, regardless of the treatment or control administered. To conclude, IL-4 and IL-1ra showed promise when administered prophylactically and improved growth and abrogated disease under conditions of App challenge. However when IL-4 and IL-1ra where administered therapeutically they did not perform as well. Moreover these compounds have potential as a commercial application to reduce the growth reduction caused by disease such as App.
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Kanzenbach, Solveig. "Wirksamkeit und Wirtschaftlichkeit einer Einstallungsmetaphylaxe in Actinobacillus-leuropneumoniae-Problembeständen mit Tulathromycin per Einmalinjektion." Berlin mbv, Mensch-und-Buch-Verl, 2010. http://d-nb.info/1002924294/04.
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Pelletier, Dora Maria. "Hemoglobin binding protein from «Actinobacillus pleuropneumoniae»: a novel method for extraction and isolation." Thesis, McGill University, 2007. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=18421.
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Membrane proteins (MPs) are the coveted, yet elusive, targets of structural genomics, structural proteomics, and the pharmaceutical industry. Characterized by amphiphilic surfaces, and lipid stabilized in vivo, MPs require unique combinations of detergent and additives for solubility and stability in vitro. We sought to bypass the exhaustive, and often unsuccessful exploration of detergents and additives for our target hemoglobin binding protein (HgbA): a 105 kDa, 22-stranded ß-barrel outer membrane protein (OMP) from Actinobacillus pleuropneumoniae. To address requirements of structural studies for milligram amounts of soluble target protein, a novel series of fractionating steps was developed to extract and isolate soluble HgbA. Two well established OMP properties were exploited in this pursuit: sarkosyl-insolubility and a robust structural architecture. Total membrane solubilization by sarkosyl detergent was modified for efficiency and fractionated insoluble OMPs from cytoplasmic membrane and soluble cytoplasmic proteins. Liberation of HgbA from the insoluble fraction required treatments more aggressive than variations of detergent and additives. Rigidified with extensive hydrogen bonding, the structural tolerance of ß-barrel proteins was exploited against elevated temperatures. Heat treatment of insoluble OMP fractions at 55 oC preferentially solubilized HgbA yielding 5mg HgbA per litre culture that retained its ability to bind hemoglobin. Protein profiles of these extraction products resolved one major band representing excess amounts of HgbA in the presence of limited quantities of minor species. This extraction protocol produces high quality HgbA that is markedly enriched from fractions that are otherwise inaccessible. Such preparations are advantageous for structural studies as well as promising for application to other ß-barrel proteins.Les protéines de la membrane externe (OMPs) sont les plus en demande pour les études structurales et pharmacologiques, mais sont en même temps les plus difficiles à travailler. Ces protéines, qui sont caractérisées par des surfaces amphiphiles et stabilisées par les lipides in vivo, requièrent des combinaisons uniques de détergents et d'additifs pour les solubiliser et les stabiliser in vitro. Pour des études structurales, nous avons voulu étudier la protéine HgbA (hemoglobin binding protein), une OMP de 105 kDa de la bactérie Actinobacillus pleuropneumoniae, comportant 22 feuillets ß antiparallèles organisés en un tonneau transmembranaire. Comme ces études demandent une grande quantité de protéine soluble, nous avons développé une nouvelle méthode d'extraction et d'isolation de HgbA par fractionnation, qui va au-delà du choix d'un détergent et d'un additif. Nous avons décidé d'exploiter deux propriétés établies des OMPs: leur insolubilité dans le détergent sarkosyl ainsi que leur structure robuste. Les OMPs ne sont pas solubilisées dans le sarkosyl et peuvent ainsi être séparées des protéines du cytoplasme et de la membrane interne. L'isolation de HgbA à partir de la fraction insoluble s'est avérée nécessiter des procédés plus agressifs que la combinaison de détergents et d'additifs. Les protéines à tonneau ß sont stabilisées par de nombreuses liaisons hydrogène, et de ce fait présentent une forte résistance structurale aux hautes températures. L'incubation à 55 oC de fractions OMP insolubles a ainsi permis de solubiliser préférentiellement HgbA, avec un rendement de 5 mg par litre de culture et une préservation de sa capacité de se lier à l'hémoglobine. La résolution de cette fraction solubilisée sur un gel SDS-PAGE montre une bande majeure, correspondant à une grande quantité de HgbA, ainsi qu'un nombre faible d'espèces mineures. Ce protocole d'extraction permet un fort enrichissement en
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Assavacheep, Pornchalit. "Environmental survival, role of exoprotein ApxIV and live vaccination studies with Actinobacillus pleuropneumoniae." Thesis, Royal Veterinary College (University of London), 2008. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.502690.
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Santos, Lucas Fernando dos. "Estudo filogenético do gene apxIA de Actinobacillus pleuropneumoniae sorotipo 5 isolados no Brasil." Universidade Federal de Viçosa, 2011. http://locus.ufv.br/handle/123456789/5063.
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Previous issue date: 2011-04-29Fundação de Amparo a Pesquisa do Estado de Minas Gerais
The toxins produced by Actinobacillus pleuropneumoniae (App) are recognized as major virulence factors found in this pathogen. Apx toxin I has strong hemolytic and cytotoxic activity, and this toxin has a high cytotoxic activity against pulmonary macrophages and neutrophils in swine. This exotoxin is produced by different serotypes, including the serotypes 1, 5, 9, 10, 11 and 14, which are involved with the most severe reported clinical cases. In general, these toxins are encoded by operons consisting of four genes: C, A, B and D, where the gene A encoding the structural proteins. The molecular basis of evolution and genetic diversity among the serotypes of App are not well understood. One of the possible explanations for this can be unavailability of DNA sequences of this bacteria in public databases. Thus, this study aimed to sequence the gene apxIA and assess the genetic diversity of this gene in isolates of the App with reference sequences available in GenBank (NCBI). In this study, we analyzed 42 isolates of serotype App 5A and 5B. The sequences of these isolates were compared with other sequences available in GenBank databases using the program MUSCLE 3.8.31 and polymorphisms of the nucleotide and amino acid sequences were analyzed. The nucleotide substitution model used was F81 + I + G, estimated from the set of sequences aligned using the program jModeltest 0.1.1. The results showed the presence of differences in the ApxI sequences that allowed group the 41 Brazilian isolates in 14 haplotypes (groups of sequences with 100% identity). Brazilian isolates were grouped into two groups (1 and 2), being distinguished by mutations in the amino acid residues 2 and 3 of protein ApxI. These results indicate the existence of a genetic diversity among isolates of App and can assist in the development of more efficient vaccine candidates.
As toxinas produzidas por Actinobacillus pleuropneumoniae (App) são, reconhecidas como os principais fatores de virulência encontrados neste patógeno. A toxina Apx I apresenta forte atividade hemolítica e citotóxica sendo considerada a exotoxina que apresenta maior atividade citotóxica contra macrófagos pulmonares e neutrófilos em suínos. Essa exotoxina é produzida por diferentes sorotipos de App, 1, 5, 9, 10, 11 e 14, sendo estes envolvidos com a maioria dos casos clínicos graves reportados. Em geral, essas toxinas são codificadas por operons constituídos por quatro genes: C, A, B e D, onde o gene A codifica a proteína estrutural. As bases moleculares da evolução e a diversidade genética existente entre os sorotipos de App ainda não estão bem compreendidas. Uma das possiveis explicações para esse fato pode ser pela reduzida disponibilidade de seqüências gênicas dessa bactéria em bancos de dados públicos. Neste sentido, este trabalho teve como objetivo sequenciar o gene apxIA e avaliar a diversidade genética desse gene de isolados brasileiros de App com as sequências de referência disponiveis no banco de dados Genbank (NCBI). Neste trabalho, foram analisados 42 isolados brasileiros de App sorotipo 5A e 5B. As sequências destes isolados foram comparadas a outras seqüências disponíveis nos bancos de dados GenBank utilizando o programa MUSCLE 3.8.31 e os polimorfismos das sequências de nucleotídeos e aminoácidos foram analisados. O modelo de substituição de nucleotídeos empregado foi o F81+I+G, estimado a partir do conjunto de sequências alinhadas utilizando o programa jModeltest 0.1.1 . Os resultados evidenciaram a presença de polimorfismos nas seqüências brasileiras que permitiram agrupar os 41 isolados, em 14 haplótipos (grupos de sequências com 100% de identidade) diferentes. Os isolados brasileiros foram agrupados em dois grupos (1 e 2), sendo distinguidos por mutações nos resíduos de aminoácidos 2 e 3 da proteína ApxI. Esses resultados indicam a existência de uma diversidade gênica entre os isolados brasileiros de App e auxiliarão no desenvolvimento de candidatos vacinais mais eficientes.
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Lavallée, Annet. "Identification et caractérisation d'un locus de fimbriae de type IV chez Actinobacillus pleuropneumoniae /." Thèse, Trois-Rivières : Université du Québec à Trois-Rivières, 2007. http://www.uqtr.ca/biblio/notice/resume/30024847R.pdf.
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Lavallée, Annet. "Identification et caractérisation d'un locus de fimbriae de type IV chez Actinobacillus pleuropneumoniae." Thèse, Université du Québec à Trois-Rivières, 2007. http://depot-e.uqtr.ca/1506/1/030024847.pdf.
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Prado, Isabelle Gonçalves de Oliveira. "Análise genômica e determinação do proteoma referência de isolados clínicos de Actinobacillus pleuropneumoniae sorotipo 8." Universidade Federal de Viçosa, 2016. http://www.locus.ufv.br/handle/123456789/10056.
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Conselho Nacional de Desenvolvimento Científico e Tecnológico
Actinobacillus pleuropneumoniae é o agente etiológico da pleuropneumonia suína, uma doença respiratória severa que resulta em significantes perdas econômicas no setor da suinocultura. Atualmente, A. pleuropneumoniae é classificado em 16 sorotipos diferentes que podem causar a doença com virulência distinta entre eles. Em Minas Gerais, Brasil, A. pleuropneumoniae é encontrado nas granjas e a maior distribuição é do sorotipo 8, sendo este até pouco tempo negligenciado em estudos de epidemiologia devido a problemas de identificação. Da mesma forma, atualmente é aceito que este sorotipo é também o de maior distribuição nos Estados Unidos, Canadá, Reino Unido além do Brasil. Até recentemente, não havia disponibilidade de genomas de A. pleuropneumoniae sorotipo 8 em banco de dados, por isso não existem informações genômicas específicas para este sorotipo, especialmente provenientes de isolados clínicos recentemente circulantes nas granjas. Neste contexto, somente a partir de 2015 os primeiros genomas foram disponibilizados para serem analisados, sendo assim sete genomas de isolados clínicos de A. pleuropneumoniae sorotipo 8 estão hoje disponíveis e estes foram analisados neste estudo. A partir das sequências genômicas dos isolados clínicos de A. pleuropneumoniae sorotipo 8 foi proposto um modelo de proteoma referência desse sorotipo com um total de 2.396 proteínas categorizadas nas diferentes classes de grupos de ortólogos. Na análise comparativa realizada entre genomas de A. pleuropneumoniae sorotipo 8 e demais sorotipos foi observado um padrão de conservação na organização do genoma entre esses. No entanto, foram identificadas algumas diferenças pontuais e dois segmentos genômicos diferenciais entre os genomas analisados com rearranjos e /ou inversões de 42,2 Kb e 59,6 Kb. O primeiro segmento foi encontrado nos genomas dos sorotipos 5 (L20), 7 (AP76) e em quatro dos isolados clínicos do sorotipo 8 investigados, sendo eles MV518, MV780, MV1022 e MV5651 contendo sequências de profago. O segundo segmento diferencial foi identificado apenas no genoma de A. pleuropneumoniae sorotipo 7 (AP76) e contém sequências como transposases caracterizando a presença de uma sequência de inserção no segmento. De acordo com a análise dos códons preferenciais de todos os sorotipos investigados, não foram observadas diferenças marcantes entre eles. Na análise comparativa dos proteomas, a maioria das sequências do proteoma referência de A. pleuropneumoniae sorotipo 8 que apresentaram baixa similaridade com os demais sorotipos foram caracterizadas como hipotéticas, não tendo sua função conhecida. Nessa porção diferencial específica foram relatadas sequências codificadoras relacionadas à plasmídeos o que caracteriza possíveis eventos de transferência horizontal de genes (THG) ao longo da história evolutiva do genoma e fatores de resistência a antibióticos como cloranfenicol, sulfonamida e tetraciclina. Na análise de similaridade entre os proteomas houve maior similaridade das proteínas do proteoma referência do sorotipo 8 com as proteínas do sorotipo 6, o que pode estar associado à dificuldade em separar esses sorotipos nas técnicas de sorotipagem. Com as análises comparativas e a caracterização das diferenças entre os sorotipos e isolados será possível compreender os mecanismos de virulência dos isolados de A. pleuropneumoniae e identificar potenciais candidatos vacinais mais eficientes.
Actinobacillus pleurapneumoniae is the causative agent of porcine pleuropneumonia, a severe respiratory illness that it results in significant economic losses in the swine industry. Currently, A. pleuropneumoniae is classified into 16 different serotypes that it can cause disease with distinct virulence between them. In Minas Gerais, Brazil, A. pleuropneumoniae is found on the farms and the largest distribution is of serotype 8, which, until shortly time, it was neglected in epidemiological studies because of the identification problems. Similarly, it is currently accepted that this serotype is also the most widely distributed in the United States of America, Canada, the United Kingdom as well as Brazil. Until recently, there was no availability of genomes of A. pleuropneumoniae serotype 8 in the database, because of this, there are no specific genomic information for this serotype, especially from of the clinical isolates that, recently, it are circulating on the farms. In this context, only from 2015, the first genomes were available to be analysed, so, today there are seven genomes of clinical isolates of A. pleuropneumoniae serotype 8 available and these were analyzed in this study. From the genomic sequences of clinical isolates of A. pleuropneumoniae serotype 8 was proposed a model of proteome which it is reference for this serotype with a total of 2.396 proteins categorized into different classes of orthologous groups. In a comparative analysis of genomes of A. pleuropneumoniae serotype 8 and other serotypes, it was observed a pattern of conservation in the organization of the genome between these. However, it were identified some specific differences and two genomic segments differentials between the genomes analyzed with rearrangements and/or reversals of 42.2 Kb and 59.6 Kb. The first segment was found in the genome of the serotype 5 (L20), 7 (AP76) and in four of the clinical isolates of serotype 8 that it were investigated: MV518, MV780, MV1022 and MV5651, which it containing prophage sequences. The second differential segment was identified only in the genome of A. pleuropneumoniae serotype 7 (AP76) and it contains sequences as transposases that it characterizes the presence of an insertion sequence in the segment. Regarding the use and the preference of codons between the serotypes investigated, there were no significant differences among them. In the comparative analysis of proteomes, the sequences of the reference proteome of A. pleuropneumoniae serotype 8, which it showed low similarity with the remaining serotypes, it were identified as hypothetical, it does not having its known function. In this specific differential portion, coding sequences associated to plasmids were described, that it characterize possible events by horizontal transfers of genes (HTG) throughout the evolutionary history of the genome and antibiotic resistance factors, such as chloramphenicol, tetracycline and sulfonamide. In the similarity analysis between the proteomes, there was greater similarity of the proteome reference of serotype 8 with the proteins of the serotype 6, which it may be associated on difficulty to separate these serotypes in the serotyping techniques. With the comparative analysis and the characterization of the differences between the serotypes and the isolates, it will be possible to understand the virulence mechanisms of the isolates of A. pleuropneumoniae and to identify vaccine candidate potential more effective.
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Medrano, Muñoz Andrés. "Expresión recombinante en E. Coli de antígenos de Actinobacillus Pleuropneumoniae para vacunación y diagnóstico." Doctoral thesis, Universitat Autònoma de Barcelona, 2003. http://hdl.handle.net/10803/3493.
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Actinobacillus pleuropneumoniae es una bacteria gramnegativa que provoca la pleuroneumonía porcina. En este trabajo se ha procedido a la producción y purificación, mediante técnicas de biología molecular, de antígenos proteicos de esta bacteria y a su uso en la formulación de una vacuna por subunidades y de un ELISA para diagnóstico. Los cuatro antígenos escogidos fueron dos proteínas de membrana externa (Tbp1 y Tbp2) y dos exotoxinas (ApxI y ApxIII).La primera necesidad consistía en localizar y secuenciar el gen tbp1 en el genoma de A. pleuropneumoniae. Para la detección de tbp1 se clonó el gen tbp2, situado en posición 5' respecto del gen tbp1, y se utilizó como sonda. Tras su identificación, el gen tbp1 fue clonado en el vector pUC119 y se obtuvo su secuencia completa, ésta se depositó en el Genbank con el código de acceso Z49708, habiéndose obtenido su patente europea EP0733708 y americana 08/624655. Finalmente, el resultado de este primer apartado fue publicado (Daban et al., 1996; Medrano et al., 1997) (Anexos VII.1 y VII.2).
Por otro lado, para la clonación de los genes apxIa y apxIIIa se utilizó una estrategia diferente. Al estar publicadas sus secuencias, se diseñaron cebadores de PCR en los cuales se introdujeron mutaciones que creaban dianas de restricción, permitiendo así la amplificación sobre el genoma y posterior clonación en vectores de expresión.
Una vez clonados los genes codificantes para las cuatro proteínas se pasó a la producción heteróloga en Escherichia coli. Para ello se utilizaron diversos vectores: pMAL, que generan como producto de expresión una proteína de fusión con la MBP (maltose binding protein) y vectores de la gama pET, con los cuales se ha desarrollado la mayor parte del trabajo. En ellos se han clonado y producido los cuatro antígenos en diversas construcciones, tanto las proteínas completas como péptidos de las mismas. A partir de los extractos obtenidos se procedió a la purificación de los antígenos por cromatografía de afinidad aprovechando fusiones con colas de histidinas.
Tras producir y purificar los cuatro antígenos (ApxI, ApxIII, Tbp1 y Tbp2) a escala de laboratorio, se procedió a la elaboración de una vacuna experimental. Se desarrollaron tres protocolos de vacunación sobre cerdos libres de anticuerpos frente a A. pleuropneumoniae. Se estudió la inocuidad y efectividad de esta vacuna.
Se determinó la respuesta humoral a partir de las muestras de suero obtenidas en los ensayos de vacunación mediante técnicas de transferencia Western-blot y de ELISA indirecto. Utilizando estas muestras y otras baterías de sueros de campo de animales con historias clínicas diversas se ha desarrollado un kit de ELISA indirecto para la detección de anticuerpos frente a A. pleuropneumoniae. Los antígenos que se utilizan para la preparación de este ELISA son la Tbp2 y la ApxI producidas de forma recombinante. Este kit se comercializa actualmente bajo el nombre de CIVTEST SUISAPP.
Actinobacillus pleuropneumoniae is a gramnegative bacteria responsible for the porcine pleuropneumonia. In this work we have proceeded to the production and purification, using molecular biology techniques, of proteinaceous antigens from this bacteria and to its use in the formulation of a subunit vaccine and in an ELISA for the serological diagnostic. The four chosen antigens were two outer membrane proteins (Tbp1 y Tbp2) and two exotoxins (ApxI y ApxIII).
The first requisite was the localization and sequenciation of the tbp1gene in the A. pleuropneumoniae genome. The tbp2 gene was cloned to be used as a probe for the tbp1 detection, this gene is located upstream the tbp1. After its location, the tbp1 gene was cloned in the pUC119 vector and his complete sequence was obtained, this sequence was submitted to the Genbank con el accession code Z49708, the European patent number is EP0733708 and the American is 08/624655. These results were published (Daban et al., 1996; Medrano et al., 1997) (Anexos VII.1 y VII.2).
On the other side, for the ApxIa y ApxIIIa genes cloning a different strategy was employed. As their sequences were already published, PCR oligonucleotides were prepared with mutations inserted in order to generate restriction sites, allowing the amplification using the genome as template followed by cloning in expression vectors.
Once the four proteins coding genes cloned we proceeded to their heterologue production in E. coli. Diverse vectors were used to achieve this: pMAL, which yields a fusion protein with the MBP (Maltose Binding Protein) and pET range vectors, most of the work has been done with the late ones. The four recombinant antigens have been cloned and produced in the pET vectors, the whole antigens and also as peptides. The recombinant proteins were purified from the expression cultures by means of affinity chromatography making use of histidine fusion tags.
After the expression and purification of the four antigens (ApxI, ApxIII, Tbp1 y Tbp2) on a laboratory scale, an experimental vaccine was prepared. This vaccine was tested in three experimental protocols using pigs free form antibodies against A. pleuropneumoniae. The harmlessness and immunogenicity of this vaccine were evaluated.
Western-blot and ELISA were used to make a serological titration on samples obtained from the vaccination assays. Using these samples and other field sera collections obtained from swine with known diverse clinical reports, an indirect ELISA assay kit was developed for the detection of specific antibodies against A. pleuropneumoniae. The antigens used in the preparation of the assay are the recombinant proteins Tbp2 and ApxI. This assay is currently being commercialised under the brand CIVTEST SUISAPP.
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Bosse, Janine T. "The contribution of urease activity to the pathogenesis of Actinobacillus pleuropneumoniae infection in pigs." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 2000. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape2/PQDD_0028/NQ51034.pdf.
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Perreault, Nancy. "Recherche de protéines exportées chez la bactérie Actinobacillus pleuropneumoniae : identification d'un locus de fimbriae /." Trois-Rivières : Université du Québec à Trois-Rivières, 2003. http://www.uqtr.ca/biblio/notice/resume/24613499R.pdf.
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Després, Kim. "Construction d'un transposon de type Tn5 pour la mutagénèse chez la bactérie Actinobacillus pleuropneumoniae /." Trois-Rivières : Université du Québec à Trois-Rivières, 2006. http://www.uqtr.ca/biblio/notice/resume/24811765R.pdf.
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Leanse, Leon. "Identification and confirmation of essential genes of Actinobacillus pleuropneumoniae in vitro and in vivo." Thesis, Imperial College London, 2017. http://hdl.handle.net/10044/1/50185.
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Actinobacillus pleuropneumoniae (APP) is a bacterial pathogen that infects the porcine respiratory tract, causing the economically significant disease porcine pleuropneumonia. As yet, no highly efficacious or heterologous vaccine has been produced for its prevention. A better understanding of host-pathogen interactions, including the contribution of essential genes is important to elucidate novel strategies for the development of both vaccines and therapeutics. This study looked to identify the essential genes for growth of APP under numerous biologically important conditions using Transposon Directed Insertion Sequencing (TraDIS) and an individual mutant screen. The loss of diversity of an original 43,000 Tn mutant library, meant TraDIS was unsuccessful, however, a total of 16 putative essential genes were identified while screening mutants anaerobically. Using RNA-seq, changes in the APP transcriptome during in vitro and in vivo conditions were also elucidated. Findings illustrated that, compared to published in vivo (acute pig infection) transcriptome data, all conditions (growth in porcine serum, anaerobically in rich medium and in the Waxmoth Galleria mellonella) showed comparable gene expression levels, suggesting their potential as alternative models for the study of APP pathogenesis. Currently, no inducible gene expression system in APP has been published. Therefore, an anhydrotetracycline (aTc) inducible gene expression system was developed to verify predicted essential genes. After initial developmental issues, the aTc-inducible system proved functional and titratable in APP strain MIDG2331 and was used to verify atpB as essential for anaerobic growth. Genes identified as essential for anaerobic growth (atpB) or highly repressed in G. mellonella (lrp, MIDG2331_00346) were evaluated for their contribution to virulence and vaccine potential in G. mellonella. The atpB and lrp mutants were attenuated for virulence in G. mellonella, whereas deletion of a gene (MIDG2331_00346), encoding a hypothetical protein, had no effect on virulence. In addition, testing of the MIDG2331 lrp mutant suggested it may be a potential live- attenuated vaccine (LAV) candidate. Therefore, using these alternative models, coupled with the use of high-throughput methods, can provide suitable alternatives to the pig for the discovery of vaccine targets.
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García, Parreño Omar Ricardo. "Persistencia de la inmunidad pasiva contra actinobacillus pleuropneumoniae en porcinos en etapa de recría." Bachelor's thesis, Universidad Nacional Mayor de San Marcos, 2007. https://hdl.handle.net/20.500.12672/7229.
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Observa la persistencia de la inmunidad pasiva en porcinos procedentes de madres seropositivas a Actinobacillus pleuropneumoniae hasta el final del período de recría en una granja tecnificada de Ica. Se utilizaron para este estudio 30 lechones los cuales fueron muestreados a los 17, 42 y 73 días, el cual corresponde al período de destete, mitad y final del período de recría respectivamente. Las muestras de suero fueron analizadas por medio de una prueba de ELISA indirecto que detecta anticuerpos contra la toxina ApxIV presente en todos los serotipos de A. pleuropneumoniae siendo ésta específica de especie. Al final de la etapa de lactación, estos animales presentaron desuniformidad en los niveles de anticuerpos, descendiendo estos niveles conforme avanzaba la edad. Al final de la fase de recría, la media del nivel de concentración de anticuerpos, expresado como el cociente de densidad óptica, resultó en 0.38, el cual indica que estos animales llegan al final de ésta etapa con niveles mínimos de anticuerpos, y se pueden volver susceptibles al ingreso de la etapa de engorde, sumado esto a factores ambientales y de estrés comunes durante ésta etapa.Tesis
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Nussbaumer, Iwan. "Zur Seroprävalenz von "Actinobacillus pleuropneumoniae" in schweizerischen Schweinezuchtbetrieben - eine Suche mit dem ApxIV ELISA /." Bern : [s.n.], 2007. http://www.ub.unibe.ch/content/bibliotheken_sammlungen/sondersammlungen/dissen_bestellformular/index_ger.html.
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Holloway, Mark-Anthony. "Genetic analysis of [Beta]-NAD-dependent and [Beta]-NAD-independent strains of Actinobacillus Pleuropneumoniae /." [S.l.] : [s.n.], 1994. http://www.ub.unibe.ch/content/bibliotheken_sammlungen/sondersammlungen/dissen_bestellformular/index_ger.html.
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Arteaga, Blanco Luis Andres. "Differential cellular immune response of hemocyte of Galleria mellonella larvae against Actinobacillus pleuropneumoniae strains." Universidade Federal de Viçosa, 2016. http://www.locus.ufv.br/handle/123456789/9267.
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Os insetos respondem à infecção através da montagem de reações imunes do tipo celular e humoral. Os reguladores primários dessas respostas são células chamadas hemócitos, os quais medeiam importantes respostas celulares, incluindo a fagocitose, encapsulamento, nodulação, e também segregam fatores humorais, tais como opsoninas, fatores de melanização e peptídeos antimicrobianos. Os hemócitos circulam ao longo da hemocele (cavidade corporal do inseto) pelo fluxo rápido da hemolinfa (sangue), além disso, partes desses hemócitos também existem como células sésseis que estão associados aos tecidos. As larvas de Galleria mellonella são uma alternativa viável para os modelos tradicionais dos mamíferos para estudar a eficácia de drogas antimicrobianas e a patogênese de microrganismos in vivo. No entanto, apesar da sua importância como um modelo de infecção, aspectos biológicos sobre as células do sistema imunológico, tais como a densidade e dinâmica dos hemócitos das larvas são mal compreendidos. No presente trabalho, investigamos a resposta imune celular dos hemócitos circulantes das larvas de G. mellonella contra diferentes cepas da bactéria Gram-negativa Actinobacillus pleuropneumoniae: baixa virulência (780), alta virulência (1022), e cepas de referência do sorotipo 8 (R8). Os hemócitos foram classificados com base no seu tamanho, morfologia, coloração e seus papeis na resposta imune, incluindo cinco tipos: prohemócitos, plasmatócitos, granulócitos, esferulócitos, e oenocitóides. Contagem total de hemócitos, contagem diferencial de hemócitos, atividade dos fagolisossomos, resposta autofágica, viabilidade celular, e a ativação da caspase-3 (como indicador de apoptose) foram determinados em hemócitos circulantes provenientes de larvas desafiadas e controle. Demostramos pela primeira vez no modelo de G. mellonella que os plasmatócitos e granulócitos ativam suas respostas autofágicas através da formação dos autofagossomos após o contato com A. pleuropneumoniae. Além disso, nossos dados demonstram que a imunidade celular do presente modelo de infecção muda dependendo do grau de virulência das cepas bacterianas.
Insects respond to infection by mounting cellular and humoral immune reactions. The primary regulators of these immune responses are cells called hemocytes, which mediate important cellular immune responses including phagocytosis, encapsulation, nodulation and also secrete immune factors such as opsonins, melanization factors and antimicrobial peptides. Hemocytes circulate through the hemocoel (body cavity) by the swift flow of hemolymph (blood), and part of these hemocytes population are sessile and are attached to tissues. Larvae of Galleria mellonella is a widely used factitious host as a viable alternative to traditional mammalian models to study the efficacy of antimicrobial drugs and the microbial pathogenesis in vivo. However, despite their importance as an infection model, biological aspects about the immune cells, such as density and hemocyte dynamic of larvae are poorly understood. In the present study, we investigated the cellular immune response of hemocytes from G. mellonella larvae against three strains of the gram-negative bacterium Actinobacillus pleuropneumoniae: low virulent (780), high virulent (1022), and the serotype 8 reference strain (R8). Five types of larval hemocytes, prohemocytes, plasmatocytes, granulocytes, oenocytoids, and spherulocytes, were distinguished according to size, morphology, detection by molecular probes, dye-staining properties, and their role in the immune response. Total hemocyte count, differential hemocyte count, lysosome activity, autophagic response, cell viability, and caspase-3 activation were determined in circulating hemocytes of naïve and infected larvae. Granulocytes and plasmatocytes were the major hemocyte types involved in the cellular defense against A. pleuropneumoniae; these hemocytes activated phagolysosome activities associated with an autophagic response against the bacteria. Moreover, our results showed that apoptosis in circulating hemocytes after exposure to virulent bacterial strains was related to an excessive autophagic cell death response induced by stress and subsequent caspase-3 activation.
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Rodrigues, Fábio Assad Féres. "Avaliação da atividade antibacteriana e antibiofilme in vitro de óleos essenciais em Actinobacillus pleuropneumoniae." Universidade Federal de Viçosa, 2017. http://www.locus.ufv.br/handle/123456789/16537.
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A antibioticoterapia é a forma de tratamento mais utilizada para o combate de doenças causadas por bactérias, tanto na medicina humana quanto na veterinária. Entretanto, o uso indiscriminado desses compostos favorece a seleção de bactérias resistentes. Uma alternativa no combate a estes patógenos é a utilização de óleos essenciais. Estes são compostos sintetizados pelas plantas a partir do metabolismo secundário e desempenham funções de atração de polinizadores e alelopatia. Neste trabalho, foi analisado o efeito antimicrobiano e antibiofilme de óleos essenciais contra isolados da bactéria Actinobacillus pleuropneumoniae, agente causal da pleuropneumonia suína. Essa doença é de grande importância na suinocultura devido às significativas perdas econômicas. Recentemente, foi observado que isolados desta bactéria apresentam genes de resistência a diversos antibióticos comerciais, tornando a prospecção e desenvolvimento de novos fármacos uma medida imediata para melhor controle da doença. Para isso, dezoito óleos essenciais foram utilizados para triagem de atividade antimicrobiana e antibiofilme em quatro isolados de A. pleuropneumoniae sorotipo 8 (MV518, MV780, MV1022 e MIDG2331) e em um isolado do sorotipo 1 (S1). Oito óleos essenciais apresentaram atividade antibacteriana, sendo, então, selecionados: canela, coentro, hortelã pimenta, hortelã do campo, tomilho, manjerona, eucalipto e louro. Estes óleos foram analisados por cromatografia gasosa para avaliar seus componentes. Os testes de concentração inibitória mínima (CIM) e de concentração bactericida mínima (CBM) foram realizados com os oito óleos ativos. Os valores de CIM e CBM foram idênticos para todos os óleos em todos os isolados, com valores variando de 5 mg/mL a 0,3125 mg/mL. O óleo de coentro (0,3125 mg/mL) e o de canela (0,625 mg/mL) apresentaram valores de CIM e CBM mais baixas para a maioria dos isolados testados, sendo considerados os mais efetivos. Adicionalmente, foram realizadas as análises de rompimento do biofilme pré-formado e da inibição do biofilme em formação. Seis óleos, na maior concentração (4xCIM), foram capazes de romper em mais de 30 % o biofilme pré-formado das bactérias analisadas, sendo que os óleos de canela e eucalipto em sua menor concentração analisada (1/8xCIM), foram capazes de romper o biofilme do isolado MV1022 em 21,29 % e 30,68 %, respectivamente. Na avaliação do biofilme em formação, cinco óleos foram capazes de inibi-lo em mais de 30 %. Entretanto, apesar do efeito no rompimento e formação do biofilme, os óleos de tomilho e coentro, dependendo da concentração utilizada, induziram o aumento na formação do biofilme. Esses resultados evidenciam a possível utilização destes óleos essenciais como sanitizantes e no combate a patógenos bacterianos. Assim, esses óleos se tornam uma possível alternativa no tratamento a doenças causadas por bactérias resistentes, a exemplo da A. pleuropneumoniae.
Antibiotic therapy is the most used form of treatment against diseases caused by bacteria, both in human and veterinary medicine. However, the indiscriminate use of these substances causes the selection of resistant bacteria. An alternative strategy of treatment against these pathogens is the utilization of essential oils. These are synthesized by aromatic plants from the secondary metabolism and perform functions of the attraction of pollinators and allelopathy. In this work, we analyzed the antimicrobial and antibiofilm effect of essential oils in isolates of Actinobacillus pleuropneumoniae, causative agent of swine pleuropneumonia. This disease is of great importance in swine farming due to significant economic losses. Recently, it was observed that isolates of this bacterium present resistance genes to several commercial antibiotics, which makes the need for prospecting and development of new drugs an immediate measure for better control of the disease. For this, eighteen essential oils were initially used for screening antimicrobial and antibiofilm activity in four isolates of A. pleuropneumoniae serotype 8 (MV518, MV780, MV1022 and MIDG2331) and a serotype 1 (S1) isolate. Eight essential oils showed antibacterial activity and were selected for future analyses: cinnamon, coriander, peppermint, mint, thyme, marjoram, eucalyptus, and laurel. These oils were analyzed by gas chromatography and presented antimicrobial components in their composition. The minimum inhibitory concentration (MIC) and minimum bacterial concentration (MBC) tests were performed with the eight active oils. MIC and MBC values were identical for all oils in all isolates, with values ranging from 5 mg/mL to 0.3125 mg/mL. Coriander oil (0.3125 mg / mL) and cinnamon oil (0.625 mg / mL) showed lower MIC and MBC for most of the tested isolates, being considered the most effective oil. In addition, the analyses of rupture of the preformed biofilm and the inhibition of the biofilm in formation were performed. Six oils, in the highest concentration (4xMIC), were able to disrupt in more than 30% the preformed biofilm of the analyzed bacteria, being the oils of cinnamon and eucalyptus in its lowest concentration analyzed (1/8xCIM), were able to break the biofilm of isolate MV1022 in 21.29 % and 30.68 %, respectively. In the evaluation of the biofilm in formation, five oils were able to inhibit this biofilm in more than 30 %. However, despite the effect on biofilm breakdown and formation, the thyme and coriander oils, depending on the concentration used, induced increased biofilm formation. These results show the possible use of essential oils as sanitizers and in the treatment against bacterial pathogens. Thus, these oils become a possible alternative in the treatment of diseases caused by resistant bacteria, such as A. pleuropneumoniae.
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Després, Kim. "Construction d'un transposon de type Tn5 pour la mutagénèse chez la bactérie Actinobacillus pleuropneumoniae." Thèse, Université du Québec à Trois-Rivières, 2006. http://depot-e.uqtr.ca/1274/1/000135761.pdf.
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Perreault, Nancy. "Recherche de protéines exportées chez la bactérie Actinobacillus pleuropneumoniae : identification d'un locus de fimbriae." Thèse, Université du Québec à Trois-Rivières, 2003. http://depot-e.uqtr.ca/1900/1/000130856.pdf.
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Höltig, Doris. "Vergleichende klinische Untersuchungen an Ferkeln der Rassen Deutsche Landrasse, Hampshire, Piétrain und Deutsches Edelschwein hinsichtlich unterschiedlicher Erkrankungsgrade nach einer Aerosolinfektion mit Actinobacillus pleuropneumoniae." Giessen VVB Laufersweiler, 2009. http://d-nb.info/995994196/04.
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Bercier, Philippe. "Effets d'Actinobacillus pleuropneumoniae et Streptococcus suis sur la barrière épithéliale de la trachée du porc." Master's thesis, Université Laval, 2019. http://hdl.handle.net/20.500.11794/37085.
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Actinobacillus pleuropneumoniae et Streptococcus suis sont des bactéries pathogènes entraînant des maladies porcines mortelles et des pertes économiques importantes dans l’industrie porcine mondiale. Ces bactéries sont notamment reconnues pour leur implication dans certaines maladies pulmonaires porcines, dont la pneumonie, et leurs mécanismes de pathogénicité ne sont pas parfaitement élucidés. De ce fait, nous nous sommes intéressés à l’activité de ces deux bactéries sur la barrière épithéliale de la trachée du porc, soit un modèle représentatif de la première ligne de défense des voies respiratoires supérieures du porc. Plus précisément, nous avons étudié la capacité d’A. pleuropneumoniae et de S. suis à induire une perte d’intégrité de la barrière épithéliale, à la traverser, et à induire une réponse inflammatoire. En considérant les cytokines pro-inflammatoires sécrétées par les cellules épithéliales en présence de ces bactéries pathogènes, les effets du TNF-α sur la barrière épithéliale ont également été analysés. Nous avons démontré qu’A. pleuropneumoniae, S. suis et le TNF-α sont aptes à induire des dommages importants à la barrière épithéliale et à permettre la translocation de ces bactéries pathogènes à travers cette barrière. Ces résultats suggèrent que la barrière épithéliale puisse représenter une cible thérapeutique contre les infections engendrées par les bactéries pathogènes environnantes afin de limiter leur capacité d’invasion. Pour ce faire, il serait nécessaire d’investiguer davantage les facteurs de virulence de ces bactéries contribuant à la perte d’intégrité de la barrière épithéliale des voies respiratoires supérieures du porc afin d’élaborer de potentiels thérapies ciblées inhibant leur pathogénicité
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Kanzenbach, Solveig [Verfasser]. "Wirksamkeit und Wirtschaftlichkeit einer Einstallungsmetaphylaxe in Actinobacillus pleuropneumoniae-Problembeständen mit Tulathromycin per Einmalinjektion / Solveig Kanzenbach." Berlin : Freie Universität Berlin, 2010. http://d-nb.info/1024502554/34.
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Bertsch, Natalie [Verfasser]. "Quantitative trait loci (QTL) für die Resistenz/Empfindlichkeit gegenüber Actinobacillus pleuropneumoniae beim Schwein / Natalie Bertsch." Gießen : Universitätsbibliothek, 2016. http://d-nb.info/1122017510/34.
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St-Arneault, Amélie. "Recherche de protéines exportées chez Actinobacillus pleuropneumoniae par la technologie de fusion avec la phosphatase alcaline /." Trois-Rivières : Université du Québec à Trois-Rivières, 2003. http://www.uqtr.ca/biblio/notice/resume/24312345R.pdf.
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Bhuller, Ravneet Kaur. "Comparative genome analyses to understand the population biology and virulence of the pig pathogen Actinobacillus pleuropneumoniae." Thesis, Imperial College London, 2017. http://hdl.handle.net/10044/1/59100.
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The bacterium Actinobacillus pleuropneumoniae is an important respiratory tract pathogen of pigs leading to major swine health problems and huge economic losses to farmers worldwide. It is divided into at least 16 distinct capsular serovars and has highly diverse pathogenicity. A safe vaccine that offers complete protection against all serovars has yet not reached the market. In this study, a multi-strain genomic approach was used to screen universal vaccine candidates against A. pleuropneumoniae. Roary was used to identify core genes between 221 A. pleuropneumoniae strains from serovars 1-5, 5b, 6-16, and K2:O7 and nontypeables strains. By cross-referencing with previous literature, it was identified that 34 of the A. pleuropneumoniae conserved genes are predicted to code for virulence factors. These included genes involved in cell wall biogenesis, anaerobic respiration, metabolism, secretion and ATP synthesis. Of the 34 genes, nine genes (ompW, prc, rbsB, tatB, atpG, dmsA, nrfAB, napB and ccmH) were identified to encode for surface proteins or secretory proteins. These surface or secretory proteins are potential candidates for subunit vaccine. Another 19 genes (murD, glpR, rfaC, malP, secB, dnaK, aspA, lpdA, purF, guaB, atpGH, moaA, moaE, lldD, frdA, hemA, hemB, ureABC and ureEG) were identified to encode for cytoplasmic proteins. These cytoplasmic proteins are potential candidates for live-attenuated or differentiating infected from vaccinated animals (DIVA) vaccines. Diversity between different A. pleuropneumoniae serovars was also examined by reconstructing phylogenetic trees based on core genes and accessory genes separately. The core gene tree clearly divided the isolates into two main groups. The genomic relationships between A. pleuropneumoniae strains belonging to the same serovar were also studied. Mostly, the isolates of the same serovar were found to be closely related to each other. The exceptions were serovars 6, 12, K2:O7 and nontypeables, which showed greater genetic variation. Due to the presence of genetic variation between and within serovars, the identification of universal vaccine candidates is the best way forward to develop subunit or DIVA based vaccines against A. pleuropneumoniae. The isolates of A. pleuropneumoniae were also investigated by formulat- ing a multilocus sequence typing (MLST) scheme based on partial sequences of the genes gdhA, infB, recA, mdh, frdB, atpG and gph and 32 sequence types (STs) were observed. The analysis of these STs using eBURST revealed six clonal complexes and suggested that A. pleuropneumoniae is an intermediately clonal bacterium. Both the MLST and the core-gene phylogeny results also revealed a possibility of capsule switching in A. pleuropneumoniae which has implications for whole-cell bacterin vaccines. Additionally, an evolutionary genomics approach was used to identify core genes that show evidence of recombination and positive selection in A. pleuropneumoniae. Approximately, 61% of the core genes showed strong signals for homologous recombination (q-value < 0.05). Furthermore, the selection analysis indicated that 25 genes are under significant selection pressure. Extensive functional analysis of the positively selected genes demonstrated that genes coding for products relevant to bacterial cell membrane, metabolism, transcription, secretion and transportation are prone to positive selection pressure. This information will be useful for researchers for novel drug development against A. pleuropneumoniae.
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St-Arneault, Amélie. "Recherche de protéines exportées chez Actinobacillus pleuropneumoniae par la technologie de fusion avec la phosphatase alcaline." Thèse, Université du Québec à Trois-Rivières, 2003. http://depot-e.uqtr.ca/1899/1/000129884.pdf.
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