A study of the use of multilocus enzyme electrophoresis as a typing tool in fowl cholera outbreaks

Abstract. The goal of this study was to characterize Pasteurella multocida isolated from rabbits. Five hundred and fifty-three apparently healthy rabbits were sampled for this study. Nasal swabs were collected from each rabbit for P. multocida isolation and identification. Isolates were further characterized by capsular and somatic antigens and genomic DNA fingerprinting. Thirty-nine P. multocida isolates were recovered from 553 rabbits (7%). Capsular typing was done by depolymerization of P. multocida capsule by Staphylococcus aureus hyaluronidase and by disc diffusion with mucopolysaccharidase enzymes (heparinase III, chondroitinase AC, and hyaluronidase). Thirty-one (79%) of the isolates were capsular type A, and 8 isolates (21%) had untypable (UT) capsules. The gel-diffusion precipitin test was used to determine the somatic type of P. multocida isolates. Nineteen isolates were somatic serotype 3 (49%), 12 were serotype 1 (31%), 1 was serotype 2, 2 were serotype 5, 2 were serotype 12 with a weak reaction to antiserum raised against serotype 7 (5%), and 1 was serotype 4. Two of the isolates (5%) were UT. Restriction endonuclease analysis of the DNA of the isolates revealed 7 distinct profiles by digestion with HindIII, and 12 profiles were obtained with HpaII, whereas digestion with EcoRI did not differentiate between any of the P. multocida DNA isolates studied. The DNA restriction endonuclease enzyme HpaII was found more useful for differentiating between DNA fingerprints of P. multocida rabbit isolates. However, no correlation between capsular type, somatic serotypes, and DNA fingerprints was seen in this study.

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“…1 These molecular techniques can thus be very useful in elucidating the epidemiology of pasteurellosis in rabbits as well.…”

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confidence: 99%

Evaluation ofPasteurella MultocidaIsolated from Rabbits by Capsular Typing, Somatic Serotyping, and Restriction Endonuclease Analysis

2004

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“…There has been no formal assessment of the reproducibility of MLEE as a typing tool for avian P. multocida. The finding that all 12 strains of an outbreak previously shown to be identical by two genotyping methods (REA and ribotyping) were also allocated to the same ET indicates that MLEE appears to have good reproducibility (Blackall et al, 1999). MLEE is not a technique that would suit routine laboratories.…”

Section: Multi-locus Enzyme Electrophoresismentioning

confidence: 88%

“…The REA and ribotyping work, along with the original field epidemiological investigation, were reported by Blackall et al (1995). The retrospective use of PFGE and REP-PCR was reported by Gunawardana et al (2000), and the retrospective use of MLEE was reported by Blackall et al (1999). Table 5 shows the direct comparison of all five typing techniques.…”

Section: A Direct Comparison Of Typing Methodsmentioning

confidence: 99%

Identification and typing ofPasteurella multocida: A review

Blackall¹,

Miflin²

2000

Avian Pathology

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Pasteurella multocida is an important pathogen of many avian species. This review critically examines recent developments in new-generation tests for the identification and typing of this bacterium. Two polymerase chain reaction (PCR) tests have been reported for P. multocida. Both tests show promise as diagnostic tests that could be considered for routine use. However, there have not yet been effective evaluation studies that examine the ability of these new tests to distinguish between P. multocida, both typical and atypical isolates, and the range of other P. multocida-like organisms found in avian species. One PCR, reported by Townsend et al. (Journal of Clinical Microbiology, 36, 1096-1100, has been the more fully evaluated and is the better choice, at this stage, for laboratories considering the use of PCR technology for detection of P. multocida. An important point is that the PCR tests have been validated by use on pure cultures or enrichment broths -not on direct examination of body tissues. To date, there have been five different technologies used to type avian P. multocida: restriction endonuclease analysis (REA), ribotyping, pulsed field gel electrophoresis, repetitive extragenic palindromic-PCR (REP-PCR) and multi-locus enzyme electrophoresis (MLEE). The methodology underlying these techniques is briefly explained and the performance of these techniques with regards to the typing of avian P. multocida is critically examined. For smaller laboratories that are investigating outbreaks of fowl cholera, it would appear that REA and REP-PCR are the typing methods of choice. For central reference laboratories that are considering studies of large collections of isolates, MLEE supported by two of the other methods would appear the most suitable techniques.

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“…Outer membrane and whole cell proteins were shown to be a suitable target for isolate typing (Dabo et al, 1999a;Davies et al, 2003b). Multi-locus enzyme electrophoresis was also used as a precise tool in characterisation of P. multocida (Blackall et al, 1999). DNA-based typing methods, mainly the restriction endonuclease techniques showing a similar discriminatory power (Blackall and Miflin, 2000), have already been established in detailed analysis of P. multocida isolates by many authors (Gardner et al, 1994;Bowles et al, 2000;El Tayeb et al, 2004).…”

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confidence: 99%

Characterisation and comparison of Pasteurella multocida isolated from different species in the Czech Republic: capsular PCR typing, ribotyping and dermonecrotoxin production

Jaglič¹,

Kučerová²,

Nedbalcová³

et al. 2005

Vet. Med. - Czech

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ABSTRACT:The aim of this study was to characterise and compare Pasteurella multocida isolates originating from pigs (n = 43), calves (n = 31), rabbits (n = 27), and to a lesser extent from other hosts (n = 6). A total of 107 P. multocida isolates were obtained from various locations in the Czech Republic. They were analysed by capsular PCR typing and ribotyping, and tested for the production of dermonecrotoxin. Most frequently, serogroup A isolates (n = 74) were found, followed by serogroup D (n = 25) and serogroup F (n = 8) isolates. From a total of fifteen different ribotypes (1-15) generated by restriction endonuclease MspI, four ribotypes (1, 3, 4, and 7) were predominant. The prevalence of predominant ribotypes was different in isolates originating from different hosts. Ribotype 1 was characteristic for rabbit isolates, ribotype 3 was primarily found in pig isolates, and ribotype 7 dominated among calf isolates. Sixteen (mainly porcine) isolates produced dermonecrotoxin but significant correlation among ribotypes and dermonecrotoxin production was not observed.

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A study of the use of multilocus enzyme electrophoresis as a typing tool in fowl cholera outbreaks

Cited by 10 publications

References 8 publications

Evaluation ofPasteurella MultocidaIsolated from Rabbits by Capsular Typing, Somatic Serotyping, and Restriction Endonuclease Analysis

Evaluation ofPasteurella MultocidaIsolated from Rabbits by Capsular Typing, Somatic Serotyping, and Restriction Endonuclease Analysis

Identification and typing ofPasteurella multocida: A review

Characterisation and comparison of Pasteurella multocida isolated from different species in the Czech Republic: capsular PCR typing, ribotyping and dermonecrotoxin production

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