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

Mbuthia, P.G.; Christensen, Henrik; Boye, Mette; Petersen, Kamille Majken Dumong; Bisgaard, Magne; Nyaga, P.N.; Olsen, John Elmerdahl

doi:10.1128/jcm.39.7.2627-2633.2001

Cited by 24 publications

(17 citation statements)

References 43 publications

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“…The oligonucleotides labeled with Cy3 and Cy5 used in the present study gave a clear bright signal, which enabled the use of narrow-band-pass filters, enhancing the signal-to-noise ratio considerably compared to those obtained with classical fluorophores, such as fluorescein and rhodamine derivatives, confirming the observations of Wessendorf and Brelje (36). However, some counterstaining can be useful for visualization of the spatial distribution of bacteria within tissue (10,22), which in the present study was achieved by the use of a double filter set. The double filter set allowed the passage of signals from the labeling dye as well as specified wavelengths of autofluorescence from the host tissue.…”

Section: Discussionsupporting

confidence: 72%

“…FISH has been applied to the sensitive detection of microorganisms in situ and has been used to reveal bacterium-host interactions at the cellular level (4). The method has proved to be a valuable tool in elucidating the pathogenesis and spatial distribution of a range of different bacteria, including Pasteurella multocida in chicken and por-cine tissues (22), Haemophilus somnus in bovine lung tissue (35), and Brachyspira hyodysenteriae and Brachyspira pilosicoli in porcine intestinal tissue (10).…”

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confidence: 99%

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Detection of Gallibacterium spp. in Chickens by Fluorescent 16S rRNA In Situ Hybridization

Bojesen¹,

Nielsen

Olsen³

et al. 2003

J Clin Microbiol

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Gallibacterium has recently been included as a new genus of the family Pasteurellaceae Pohl 1981, which encompasses bacteria previously reported as Pasteurella anatis, "Actinobacillus salpingitidis," and avian Pasteurella haemolytica-like organisms. So far, identification has exclusively relied on phenotypic characterization. We present a method based on a cyanine dye 3.18-labeled in situ hybridization probe targeting 16S rRNA to allow specific detection of bacteria belonging to the genus Gallibacterium. The probe, GAN850, showed no crossreactivity to 25 other poultry-associated bacterial species, including members of the families Pasteurellaceae, Enterobacteriaceae, and Flavobacteriaceae, when cross-reactivities were evaluated by whole-cell hybridization. The probe was further evaluated by hybridization to formalin-fixed spleen and liver tissues from experimentally infected chickens, in which it proved to be useful for the detection of Gallibacterium. Additionally, determination of the spatial distribution and the host cell affiliation of Gallibacterium at various times during the infection process was possible. In conclusion, the in situ hybridization technique described may be of use as a diagnostic tool as well as for studies to elucidate the pathogenesis of Gallibacterium infections in chickens.

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Section: Discussionsupporting

confidence: 72%

mentioning

confidence: 99%

Detection of Gallibacterium spp. in Chickens by Fluorescent 16S rRNA In Situ Hybridization

Bojesen¹,

Nielsen

Olsen³

et al. 2003

J Clin Microbiol

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“…Development of specific DNA tools for the detection and identification of P. multocida has previously been hampered by non-specific PCR and in situ detection of P. avium biovar 2 and P. canis biovar 2 (Mbuthia et al, 2001;Miflin & Blackall, 2001;Townsend et al, 2001). The data presented in the present paper fully document that these organisms should be reclassified with P. multocida, since genotypically they represent genuine P. multocida.…”

Section: Discussionmentioning

confidence: 81%

“…Specific detection is also possible based on the psl gene combined with hybridization (Kasten et al, 1997). Problems with the non-specificity of PCR and in situ hybridization tests for P. multocida have recently been reported for biovar 2 of P. avium and biovar 2 of P. canis (Mbuthia et al, 2001;Miflin & Blackall, 2001;Townsend et al, 2001).…”

Section: V-factor (Nad)mentioning

confidence: 99%

Revised description and classification of atypical isolates of Pasteurella multocida from bovine lungs based on genotypic characterization to include variants previously classified as biovar 2 of Pasteurella canis and Pasteurella avium

Angen

Olsen²,

Bisgaard³

2004

Microbiology

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Strains deviating in key phenotypic characters, mainly isolated from cases of bovine pneumonia in five European countries, were genotyped in order to examine their genotypic relationship with Pasteurella multocida. Twenty-two strains of Pasteurella avium biovar 2, including variants in indole, xylose and mannitol, 18 strains of Pasteurella canis biovar 2 and variants of this taxon, five strains of P. multocida subsp. septica showing variations in indole and ornithine decarboxylase, nine strains of P. multocida subsp. multocida showing variation in ornithine decarboxylase and mannitol, and type strains of the subspecies of P. multocida were included. Ribotyping was used to examine the relationship of the strains, and 13 types, each containing between one and 20 isolates, were observed. Identical ribotypes were observed in some cases for P. avium biovar 2 and either P. canis biovar 2 or P. multocida subsp. septica. ITS (16S–23S rRNA internal transcribed spacer) fragment-length profiling showed identity of the majority of strains (47 of 52), representing all four taxa, with only five divergent strains. A 16S rRNA sequence comparison of 11 strains representing the main ribotype clusters showed 99·9 % similarity to the type strain of P. multocida subsp. multocida, but only 97·4 % similarity was obtained to P. canis (biovar 1) and 93·7 % to P. avium (biovar 1). A species-specific PCR test for P. multocida gave a positive result with biovar 2 variants of P. avium and P. canis. DNA–DNA hybridizations between strains of P. multocida, biovar 2 variants of P. avium and P. canis, and P. multocida subsp. septica confirmed similarity at the species level. It is proposed, on the basis of genotypic similarity, that P. multocida be reclassified to include the biovar 2 variants of P. avium and P. canis and that the existence of the biovar 2 variants of P. avium and P. canis is highly questionable. It is concluded that the redefined P. multocida is genotypically homogeneous, although phenotypically diverse lineages exist with respect to ornithine decarboxylase, indole and mannitol, characters that have been regarded as essential for identification to the species level. A formal reclassification of the species is not possible, however, since too few strains have been found to vary in these key characters. Considering the phenotypic diversity of P. multocida, identification will have to depend partly on genotypic methods and the source host also seems important for safe diagnosis.

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“…Serotype B:3 is often isolated from avian disease cases that manifest as sinusitis (229,230) with symptoms of nasal discharge, increased lacrimation, swelling, and inflammation. The respiratory tract appears to be the primary site of infection for fowl cholera (126,129,223,231), but isolation of P. multocida subsp. multocida from avian salpingitis has also been reported (232).…”

Section: Pasteurellosis and Fowl Choleramentioning

confidence: 99%

Pasteurella multocida: from Zoonosis to Cellular Microbiology

2013

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SUMMARY In a world where most emerging and reemerging infectious diseases are zoonotic in nature and our contacts with both domestic and wild animals abound, there is growing awareness of the potential for human acquisition of animal diseases. Like other Pasteurellaceae , Pasteurella species are highly prevalent among animal populations, where they are often found as part of the normal microbiota of the oral, nasopharyngeal, and upper respiratory tracts. Many Pasteurella species are opportunistic pathogens that can cause endemic disease and are associated increasingly with epizootic outbreaks. Zoonotic transmission to humans usually occurs through animal bites or contact with nasal secretions, with P. multocida being the most prevalent isolate observed in human infections. Here we review recent comparative genomics and molecular pathogenesis studies that have advanced our understanding of the multiple virulence mechanisms employed by Pasteurella species to establish acute and chronic infections. We also summarize efforts being explored to enhance our ability to rapidly and accurately identify and distinguish among clinical isolates and to control pasteurellosis by improved development of new vaccines and treatment regimens.

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

Cited by 24 publications

References 43 publications

Detection of Gallibacterium spp. in Chickens by Fluorescent 16S rRNA In Situ Hybridization

Detection of Gallibacterium spp. in Chickens by Fluorescent 16S rRNA In Situ Hybridization

Revised description and classification of atypical isolates of Pasteurella multocida from bovine lungs based on genotypic characterization to include variants previously classified as biovar 2 of Pasteurella canis and Pasteurella avium

Pasteurella multocida: from Zoonosis to Cellular Microbiology

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