Pasteurella multocida detection by 5′ Taq nuclease assay: A new tool for use in diagnosing fowl cholera

Corney, Bruce G.; Diallo, Ibrahim S.; Wright, Liam; Hewitson, Glen; Jong, Anja J. de; Burrell, P. C.; Duffy, P F; Stephens, C.P.; Rodwell, B.J.; Boyle, D.B.; Blackall, P. J.

doi:10.1016/j.mimet.2007.01.014

Cited by 24 publications

(19 citation statements)

References 18 publications

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“…PHA test was conducted to determine the humoral immune response of the serum of chickens having been inoculated at 9 weeks aged birds with the antigen containing P. multocida as per the method described by Chang (1987) but slight modification was done as suggested by Kuczkowski et al, (2006) and Corney et al, (2007). The pre-vaccination PHA titres of sera samples of all vaccinated and control birds was found with a mean of <4.00±0.00 that was closely related with Mondal et al (1988).…”

Section: Discussionmentioning

confidence: 99%

Isolation and Identification of Pasteurella multocida from Chicken for the Preparation of Oil Adjuvanted Vaccine

et al. 2013

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A B S T R A C TThe research work was performed for the isolation and identification of Pasteurella multocida from field cases, preparation of oil adjuvanted vaccine from isolated strain and determination of its efficacy. Samples were collected from suspected dead birds of three poultry farms of Bangladesh (Code name: M and R). The P. multocida isolates were Gram negative, non-motile, non-spore forming rod occurring singly or pairs and occasionally as chains or filaments. Biochemically P. multocida ferment basic sugar and consistently produced acid except from maltose and lactose. After isolation formalin killed oil adjuvanted Fowl cholera vaccine was prepared in Laboratory of the Department of Microbiology and Hygiene, BAU and this experimental vaccine (3.2x10 8 CFU/ml) was administered in nine weeks old White Leg Horn chickens at the different dose rate through intramuscular (IM) route in each selected group A (1ml alum precipitated vaccine), B (0.5ml alum precipitated vaccine), C (1ml oil adjuvanted vaccine) and D (0.5ml oil adjuvanted vaccine). Pre-vaccinated sera were collected from all groups of birds. The mean of Passive Hemagglutination (PHA) titers of post-vaccination were 51±17.8, 76.8±17, 89.6±17, and 115±17.81 in group A, B, C and D respectively which consist of 5 birds in each. The vaccine produced better immune response when boostering with the similar dose and route at 15 days after primary vaccination. The mean PHA titers were higher at group D than other groups after boostering. Challenge infection was conducted on all the vaccinated and control group (n=5) of birds after 15 days of vaccination which protect 93.75% of birds and the PHA titers from different groups analyzed to determine the protective capacity of vaccinated chickens against challenge exposure. It was demonstrated that experimental oil adjuvanted fowl cholera vaccine with 0.5ml dose produce higher immune response against challenge infection and found to be safe.

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Section: Discussionmentioning

confidence: 99%

Isolation and Identification of Pasteurella multocida from Chicken for the Preparation of Oil Adjuvanted Vaccine

et al. 2013

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“…Primer design was based on the 16S rRNA gene sequences of P. multocida from the GenBank database and from reference strains in the current study (Tables S1, S2). The forward primer, PMA2f (59-ATAACTGTGGGAAACTGCAGCTAA-39) was adopted from Corney et al (2007) and showed a perfect match to all sequences. This primer showed some tendency for homodimers with a free energy of melting (DG) of 210.24 kcal mol 21 with 39 overhangs and weak tendency for hairpins with DG of 22.24 kcal mol 21 (OligoAnalyzer 3.1) (Owczarzy et al, 2008).…”

Section: Methodsmentioning

confidence: 99%

“…The PCR designed to distinguish between lineages A and B is not specific for P. multocida, since sequences from other members of Pasteurellaceae were predicted to be amplified as well. For species specificity we recommend other PCRs such as those of Corney et al (2007) or Townsend et al (1998). Recently a RFLP procedure based on restriction enzymic digestion of PCR-amplified 16S rRNA was published that also allowed the separation of A and B lineages of P. multocida (Sellyei et al, 2012).…”

Section: Ipmentioning

confidence: 99%

Multilocus sequence analysis of Pasteurella multocida demonstrates a type species under development

Bisgaard

Petersen

2013

Microbiology

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The aim of the present study was to use multilocus sequence typing (MLST) of a diverse collection of Pasteurella multocida with regard to animal source, place and date of collection, including all available serovars of Carter, Heddleston, Little & Lyon, Namioka, Cornelius and Roberts, to further investigate the evolution of this species with a focus on two lineages, A (P. multocida subsp. multocida and P. multocida subsp. gallicida) and B (P. multocida subsp. septica), previously reported. Isolates of P. multocida (n5116) including reference strains of major serotyping systems were investigated by MLST based on partial sequences of the genes adk, est, gdh, mdh, pgi, pmi and zwf, and 67 sequence types (STs) were observed. Phylogenetic analysis of these concatenated sequences confirmed the separation of groups A (41 STs, 71 isolates) and B (22 STs, 38 isolates) out of the 67 STs. All Carter serovars, 12 Heddleston serovars, all three Little-Lyon types, six out of seven Namioka serovars, all five Roberts types and all four Cornelius serovars were allocated to the A group, while group B included the remaining four Heddleston serovars, 6, 7, 8 and 13, in addition to Namioka type 8 : A. The overrepresentation of reference strains of serotyping systems in the A group contrasts with the high number of isolates obtained from diseased birds in the B group, the effect of which should be addressed in future vaccine development. Isolates from birds (25) dominated the B group, which also included four isolates from Felidae, whereas group A included isolates from all types of hosts. The evolutionary implications of the lack of capsular type D, pig and bovine isolates in group B, as well as its association with Aves and Felidae that also applied to the whole Rural Industries Research and Development Corporation (RIRDC) MLST database, need further investigation. The combination of rpoB and 16S rRNA gene sequence comparison as well as the developed PCR test assigned isolates to lineage A, represented by the type strain of P. multocida subsp. Multocida, or lineage B represented by the type strain of P. multocida subsp. septica. It was not possible to circumscribe either the A or B lineages with a set of conserved phenotypic characters, calling into question the validity of subspecies within P. multocida. Phylogenetic analysis carried out on individual MLST genes showed deviations as to single or multiple genes for 17 % of group A and 43 % of group B, indicating that lineage A probably developed from lineage B, and that major changes are ongoing. From a genotypical point of view, we conclude that P. multocida subsp. gallicida represents an artificial unit.

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“…However, many other factors may be related, including genes encoding structures such as fimbriae and bacterial adhesins or outer membrane proteins (Corney et al 2007, Hatfaludi et al 2010). In addition, there are genes encoding enzymes involved in the uptake of iron, genes encoding enzymes involved in bacterial metabolism and genes that encode exotoxins (Harper et al 2006).…”

Section: Introductionmentioning

confidence: 99%

Detection of virulence-associated genes of Pasteurella multocida isolated from cases of fowl cholera by multiplex-PCR

et al. 2013

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The current systems of breeding poultry, based on high population density, increase the risk of spreading pathogens, especially those causing respiratory diseases and those that have more than one host. Fowl Cholera (FC) is one such pathogen, and even though it represents one of several avian diseases that should be considered in the differential diagnosis of notifiable diseases that present with sudden death, the pathogenesis and virulence factors involved in FC are still poorly understood. The objective of this study was to investigate twelve genes related to virulence in 25 samples of Pasteurella multocida isolated from FC cases in the southern region of Brazil through the development of multiplex PCR protocols. The protocols developed were capable of detecting all of the proposed genes. The ompH, oma87, sodC, hgbA, hgbB, exBD-tonB and nanB genes were present in 100% of the samples (25/25), the sodA and nanH genes were present in 96% (24/25), ptfA was present in 92% (23/25), and pfhA was present in 60% (15/25). Gene toxA was not identified in any of the samples studied (0/25). Five different genetic profiles were obtained, of which P1 (negative to toxA) was the most common. We concluded that the multiplex-PCR protocols could be useful tools for rapid and simultaneous detection of virulence genes. Despite the high frequency of the analyzed genes and the fact that all samples belonged to the same subspecies of P. multocida, five genetic profiles were observed, which should be confirmed in a study with a larger number of samples.

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Pasteurella multocida detection by 5′ Taq nuclease assay: A new tool for use in diagnosing fowl cholera

Cited by 24 publications

References 18 publications

Isolation and Identification of Pasteurella multocida from Chicken for the Preparation of Oil Adjuvanted Vaccine

Isolation and Identification of Pasteurella multocida from Chicken for the Preparation of Oil Adjuvanted Vaccine

Multilocus sequence analysis of Pasteurella multocida demonstrates a type species under development

Detection of virulence-associated genes of Pasteurella multocida isolated from cases of fowl cholera by multiplex-PCR

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