<i>Actinobacillus pleuropneumoniae</i> Osteomyelitis in Pigs Demonstrated by Fluorescent In Situ Hybridization

Jensen, Tim Kåre; Boye, Mette; Hagedorn-Olsen, Tine; Riising, H.-J.; Angen, Øystein

doi:10.1354/vp.36-3-258

Cited by 27 publications

(23 citation statements)

References 9 publications

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“…[29][30][31] In each test round, sections from lung tissue containing A. pleuropneumoniae, as verified by bacteriology and FISH, were included as positive staining controls.…”

Section: Fishmentioning

confidence: 99%

“…For the specific detection of A. pleuropneumoniae, sections were examined by FISH using a fluorescein isothiocyanate (FITC)-labelled oligonucleotide probe (Ap185; MWGbiotech AG, Ebersberg, Germany), with the sequence 5 0 -CCCACCCTTTAATCCATAG-3 0 , targeting 16S rRNA of A. pleuropneumoniae, as previously described. [29][30][31] In each test round, sections from lung tissue containing A. pleuropneumoniae, as verified by bacteriology and FISH, were included as positive staining controls.…”

Section: Fishmentioning

confidence: 99%

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Pulmonary infections in swine induce altered porcine surfactant protein D expression and localization to dendritic cells in bronchial‐associated lymphoid tissue

Soerensen¹,

Holmskov

Aalbæk³

et al. 2005

Immunology

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SummarySurfactant protein D (SP-D) is a pattern-recognition molecule of the innate immune system that recognizes various microbial surface-specific carbohydrate and lipid patterns. In vitro data has suggested that this binding may lead to increased microbial association with macrophages and dendritic cells. The aim of the present in vivo study was to study the expression of porcine SP-D (pSP-D) in the lung during different pulmonary bacterial infections, and the effect of the routes of infection on this expression was elucidated. Furthermore, the aim was to study the in vivo spatial relationship among pSP-D, pathogens, phagocytic cells and dendritic cells. Lung tissue was collected from experimental and natural bronchopneumonias caused by Actinobacillus pleuropneumoniae or Staphylococcus aureus, and from embolic and diffuse interstitial pneumonia, caused by Staph. aureus or Arcanobacterium pyogenes and Streptococcus suis serotype 2, respectively. By comparing normal and diseased lung tissue from the same lungs, increased diffuse pSP-D immunoreactivity was seen in the surfactant in both acute and chronic bronchopneumonias, while such increased expression of pSP-D was generally not present in the interstitial pneumonias. Co-localization of pSP-D, alveolar macrophages and bacteria was demonstrated, and pSP-D showed a patchy distribution on the membranes of alveolar macrophages. SP-D immunoreactivity was intracellular in dendritic cells. The dendritic cells were identified by their morphology, the absence of macrophage marker immunoreactivity and the presence of dendritic cell marker immunoreactivity. Increased expression of pSP-D in the surfactant coincided with presence of pSP-D-positive dendritic cells in bronchus-associated lymphoid tissue (BALT), indicating a possible transport of pSP-D through the specialized M cells overlying (BALT). In conclusion, we have shown that pSP-D expression in the lung surfactant is induced by bacterial infection by an aerogenous route rather than by a haematogenous route, and that the protein interacts specifically with alveolar macrophages and with dendritic cells in microbial-induced BALT. The function of the interaction between pSP-D and dendritic cells in BALT remain unclear, but pSP-D could represent a link between the innate and adaptive immune system, facilitating the bacterial antigen presentation by dendritic cells in BALT.

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“…[29][30][31] In each test round, sections from lung tissue containing A. pleuropneumoniae, as verified by bacteriology and FISH, were included as positive staining controls.…”

Section: Fishmentioning

confidence: 99%

Section: Fishmentioning

confidence: 99%

Pulmonary infections in swine induce altered porcine surfactant protein D expression and localization to dendritic cells in bronchial‐associated lymphoid tissue

Soerensen¹,

Holmskov

Aalbæk³

et al. 2005

Immunology

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“…FISH has been used to visualize the spatial distribution of Escherichia coli in intestinal mucosa (42); to evaluate colonization of mouse intestines by Salmonella enterica serovar Typhimurium (32); and to specifically detect Salmonella serovars in pig intestines and mouse lungs (40), Actinobacillus pleuropneumoniae strains in diseased porcine tissues (23,26), Brachyspira (Serpulina) pilosicoli infections in the intestines of growing pigs (11,27), Lawsonia intracellularis in porcine proliferative enteropathy (10), Streptococcus suis infection in pigs (12), and bacteria in blood cultures (25,29).…”

mentioning

confidence: 99%

Specific Detection of Pasteurella multocida in Chickens with Fowl Cholera and in Pig Lung Tissues Using Fluorescent rRNA In Situ Hybridization

Mbuthia

Christensen

Boye³

et al. 2001

J Clin Microbiol

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A Pasteurella multocida species-specific oligonucleotide probe, pmhyb449, targeting 16S rRNA was designed and evaluated by whole-cell hybridization against 22 selected reference strains in animal tissues. It differentiated P. multocida from other bacterial species of the families Pasteurellaceae and Enterobacteriaceae and also from divergent species of the order Cytophagales (except biovar 2 strains of Pasteurella avium and Pasteurella canis, which have high 16S rRNA similarity to P. multocida). The potential of the probe for specific identification and differentiation of P. multocida was further detected in formalin-fixed paraffin-embedded lung tissues from experimental fowl cholera in chickens and infections in pigs. In chicken lung tissues P. multocida cells were detected singly, in pairs, as microcolonies, and as massive colonies within air capillaries (septa and lumen), parabronchial septa, and blood vessels (wall and lumen). In pig lung, postmortem-injected P. multocida was detected in the alveoli (lumen and wall), and in both animals the bacterial cells were seen in the bronchi. The results showed that with the oligonucleotide probe pmhyb449, fluorescent in situ hybridization is a suitable and fast method for specific detection of P. multocida in histological formalin-fixed tissues. The test was replicable and reproducible and is recommended as a supplementary test for diagnosis and as a tool in pathogenesis studies of fowl cholera and respiratory tract infections in pigs due to P. multocida.

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“…The results of an earlier study indicate that fluorescent in situ hybridization using a DNA probe designed from the 16S ribosomal RNA of A. pleuropneumoniae can detect the 12 serotypes of A. pleuropneumoniae [9,10]. However, that study did not examine lung tissue from pigs infected with all of the A. pleuropneumoniae serotypes, and the use of fluorescent in situ hybridization did not preserve cellular and histological details.…”

Section: Discussionmentioning

confidence: 96%

In situ hybridization for the detection of the apxIV gene in the lungs of pigs experimentally infected with twelve Actinobacillus pleuropneumoniae serotypes

2002

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-The detection of the apxIV gene in lung tissues from pigs experimentally infected with the 12 major A. pleuropneumoniae serotype (1 to 12) reference strains was studied by in situ hybridization using a non-radioactive digoxigenin-labeled DNA probe. In situ hybridization produced a distinct positive signal in all pigs inoculated with the 12 A. pleuropneumoniae serotypes. Positive hybridization typically exhibited a dark-brown to black reaction product in intracellular and extracellular locations, without background staining. A strong hybridization signal was seen in degenerated alveolar leukocytes ("oat cells") adjacent to the foci of coagulative necrosis and in the alveolar spaces. The in situ hybridization methodology developed for the detection of the apxIV gene is a valuable tool for the diagnosis of porcine pleuropneumonia caused by A. pleuropneumoniae when only formalin-fixed tissues are submitted for diagnosis.Actinobacillus pleuropneumoniae / Apx toxin / in situ hybridization / lung / pig Résumé -Détection par hybridation in situ du gène apxIV dans les poumons de porcs infectés expérimentalement par douze sérotypes de Actinobacillus pleuropneumoniae. La détection du gène apxIV dans les tissus pulmonaires de porcs infectés expérimentalement par les douze sérotypes majeurs des souches de référence (1 à 12) de Actinobacillus pleuropneumoniae a été effectuée par hybridation in situ utilisant une sonde à ADN non radioactive marquée à la digoxigénine. L'hybridation in situ a produit un signal positif distinct chez tous les porcs inoculés par les douze sérotypes de A. pleuropneumoniae. Un résultat positif d'hybridation entraînait typiquement un produit de réaction marron foncé à noir localisé en intra-et en extra-cellulaire, sans bruit de fond. Un signal fort d'hybridation a été observé dans les leucocytes alvéolaires dégénérés adjacents aux foyers de nécrose et dans les espaces alvéolaires. La méthode d'hybridation in situ développée pour la détection du gène apxIV est un outil précieux pour le diagnostic de la pleuropneumonie porcine causée par A. pleuropneumoniae lorsque seuls les tissus fixés dans le formol sont soumis pour le diagnostic.Actinobacillus pleuropneumoniae / Apx toxine / hybridation in situ / poumon / porc 653 Vet.

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Actinobacillus pleuropneumoniae Osteomyelitis in Pigs Demonstrated by Fluorescent In Situ Hybridization

Cited by 27 publications

References 9 publications

Pulmonary infections in swine induce altered porcine surfactant protein D expression and localization to dendritic cells in bronchial‐associated lymphoid tissue

Pulmonary infections in swine induce altered porcine surfactant protein D expression and localization to dendritic cells in bronchial‐associated lymphoid tissue

Specific Detection of Pasteurella multocida in Chickens with Fowl Cholera and in Pig Lung Tissues Using Fluorescent rRNA In Situ Hybridization

In situ hybridization for the detection of the apxIV gene in the lungs of pigs experimentally infected with twelve Actinobacillus pleuropneumoniae serotypes

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