Molecular Epidemiology of Japanese Avian Pasteurella multocida Strains by the Single-Enzyme Amplified Fragment Length Polymorphism and Pulsed-Field Gel Electrophoresis

Abstract

ABSTRACT. Molecular epidemiology analyses of the 36 clinical isolates of Pasteurella multocida from various avian hosts in Japan between 1976 to 2007 including 5 reference strains from the U.S.A., Taiwan and Indonesia were performed by employing the single-enzyme amplified fragment length polymorphism (SE-AFLP) comparison with the classical ApaI-based pulsed-field gel electrophoresis (PFGE). As the results, SE-AFLP gave 21 profiles while PFGE gave 20 profiles. The Simpson's index of diversity analysis indicated that SE-AFLP gave a high discrimination power than PFGE. This concluded that SE-AFLP is a higher discrimination power than PFGE to differentiate avian P. multocida isolates in Japan. In addition, the genetical profiles suggested that there is the evolution of somatic serotype 3 strain in the indigenous host of Japan.KEY WORDS: discrimination power, Pasteurella multocida, PFGE, SE-AFLP.J. Vet. Med. Sci. 72(11): 1465-1470, 2010 Fowl cholera is an important disease in poultry industry around the world. Pasteurella multocida of the gram negative bacteria is recognized as the etiologic agent of fowl cholera [16,18]. The bacterium can be classified by capsular serogrouping into 5 serogroups (A, B, D, E and F) [6,17] and by lipopolysaccharide into 16 somatic serotypes (1-16) [5]. Fowl cholera usually appears a septicemic disease in the acute or peracute conditions associated with the high mortality and morbidity but the chronic conditions are also observed [16,18]. In Japan, strain of P. multocida capsular serogroup A and somatic serotype 3 was predominated among avian isolates and strains of somatic serotypes 1, 4 and 10 were also isolated [20]. Some of the clinical isolates were determined for their capsulation and virulence for chicken by the observation of their colonies by electron microscope or obliquely-transmitted light stereomicroscope and experimental infection in chickens [4]. Our previous investigations indicated the genetic variation of the adhesive protein gene among each isolates by PCR-RFLP employing the HindIII and EcoRI digestion [21]. However, the genetic epidemiology of these isolates needs to be clarified.Since a number of DNA-based typing systems have been employed to characterize the isolates [1,3,10,14,15]. Pulsed-field gel electrophoresis (PFGE) is recognized as the classical DNA-based typing method of the isolates. However, the higher discrimination power DNA-based typing methods have been demonstrated. Previous studies suggested that the amplified fragment length polymorphism (AFLP) have been recognized as the typing method that showed the higher power of discrimination than either PFGE or the random amplified polymorphism DNA (RAPD) [1,8]. However, the DNA-based typing method requires more knowledge about DNA to analyze [8]. Then, the recent study employed the SE-AFLP and PFGE to differentiate avian P. multocida isolates in Japan and the discriminatory power of the typing methods was also compared. MATERIALS AND METHODSBacterial strains and growth condition: P. multocida strains were ...

Antimicrobial susceptibility, serotypes and genotypes of Pasteurella multocida isolates associated with swine pneumonia in Taiwan

“…Previous studies used Apa I, Bsp 120I and Sma I 39 41 42 . Apa I was reported to be suitable for the phylogenetic analysis of PM isolated from a broad range of species and the results were distinguishable in serotypes and strains with different virulence 19 40 41 . The aim of this study was to examine whether a correlation between genotypes and serotypes exist, so PFGE was employed to analyse the PM strains.…”

“…38 PFGE analyses of PM isolated from poultry, swine, cattle and human infections have been conducted in many previous studies. [39][40][41] The selection of restriction enzymes varied in these stud-ies. Previous studies used ApaI, Bsp120I and SmaI.…”

“…19 In the present study, the results showed that the major strain of PM was the capsular type D. The distribution of PM serotypes in this study was also different from the aforementioned Pors and other studies, and this could be due to the different sources of sampling in this study. PM serotypes can be easily clustered using PFGE genotyping 41 and the PFGE genotyping can also distinguish PM strains isolated from humans and swine. 40 The present study demonstrated that PFGE is a useful tool to categorise the serotypes and the profiles of strains with toxins of PM.…”

Investigation of genetic diversity and epidemiological characteristics of <i>Pasteurella multocida</i> isolates from poultry in southwest China by population structure, multi-locus sequence typing and virulence-associated gene profile analysis

“…[ 14 ] simplified the typing of LPS antigens (L1-L8) and developed a multiplex PCR targeting the genes encoding the LPS structures (LPS-mPCR). Gene technologies such as 16S rRNA , restriction endonuclease analysis (REA), ribotyping, random amplification of polymorphic DNA (RAPD)-PCR, pulsed-field gel electrophoresis (PFGE) and multilocus sequence type analysis (MLST), have pushed forward studies of molecular epidemiology and genetic diversity of P. multocida [ 24 , 35 , 43 ]. MLST is considered the appropriate bacterial typing techniques for global and long-term studies [ 13 , 36 ] and the results can be easily compared among different laboratories for long-term and global surveillance of bacterial strains [ 36 , 43 ].…”