Geographically diverse Australian isolates of<i>Melissococcus pluton</i>exhibit minimal genotypic diversity by restriction endonuclease analysis

Djordjevic, Steven P.; Smith, Lisa; Forbes, Wendy Ann; Hornitzky, M.

doi:10.1111/j.1574-6968.1999.tb13519.x

Cited by 25 publications

(10 citation statements)

References 13 publications

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“…Despite their importance, our understanding of M. plutonius remains very limited. Although M. plutonius has been thought to be homologous or clonal [7]–[9], our results demonstrated that this species consists of at least two groups of strains (typical and atypical M. plutonius ) that are phenotypically and genetically distinguishable. Interestingly, irrespective of the geographical source and year of isolation, all typical M. plutonius strain/isolates showed very similar PFGE profiles.…”

Section: Discussionmentioning

confidence: 57%

“…M. plutonius is a one genus-one species bacterium and is thought to be remarkably homogeneous based on morphological, physiological, immunological, and genetic studies [7]–[9]. For instance, Djordjevic et al [9] reported that Australian M. plutonius isolates originating from geographically diverse regions were markedly similar in their whole cell proteins, immunoreactive antigens, and DNA restriction endonuclease profiles.…”

Section: Introductionmentioning

confidence: 99%

“…For instance, Djordjevic et al [9] reported that Australian M. plutonius isolates originating from geographically diverse regions were markedly similar in their whole cell proteins, immunoreactive antigens, and DNA restriction endonuclease profiles. Moreover, they showed genetic homogeneity among Australian and British isolates and suggested that this species may be clonal [9]. However, the geographical locations of the isolates characterized so far have been mainly Europe, North and South America, and Australia.…”

Section: Introductionmentioning

confidence: 99%

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Diversity of Melissococcus plutonius from Honeybee Larvae in Japan and Experimental Reproduction of European Foulbrood with Cultured Atypical Isolates

Arai

Tominaga²,

et al. 2012

PLoS ONE

154

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European foulbrood (EFB) is an important infectious disease of honeybee larvae, but its pathogenic mechanisms are still poorly understood. The causative agent, Melissococcus plutonius , is a fastidious organism, and microaerophilic to anaerobic conditions and the addition of potassium phosphate to culture media are required for growth. Although M. plutonius is believed to be remarkably homologous, in addition to M. plutonius isolates with typical cultural characteristics, M. plutonius -like organisms, with characteristics seemingly different from those of typical M. plutonius , have often been isolated from diseased larvae with clinical signs of EFB in Japan. Cultural and biochemical characterization of 14 M. plutonius and 19 M. plutonius -like strain/isolates revealed that, unlike typical M. plutonius strain/isolates, M. plutonius -like isolates were not fastidious, and the addition of potassium phosphate was not required for normal growth. Moreover, only M. plutonius -like isolates, but not typical M. plutonius strain/isolates, grew anaerobically on sodium phosphate-supplemented medium and aerobically on some potassium salt-supplemented media, were positive for β-glucosidase activity, hydrolyzed esculin, and produced acid from L-arabinose, D-cellobiose, and salicin. Despite the phenotypic differences, 16S rRNA gene sequence analysis and DNA-DNA hybridization demonstrated that M. plutonius -like organisms were taxonomically identical to M. plutonius . However, by pulsed-field gel electrophoresis analysis, these typical and atypical ( M. plutonius -like) isolates were separately grouped into two genetically distinct clusters. Although M. plutonius is known to lose virulence quickly when cultured artificially, experimental infection of representative isolates showed that atypical M. plutonius maintained the ability to cause EFB in honeybee larvae even after cultured in vitro in laboratory media. Because the rapid decrease of virulence in cultured M. plutonius was a major impediment to elucidation of the pathogenesis of EFB, atypical M. plutonius discovered in this study will be a breakthrough in EFB research.

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Section: Discussionmentioning

confidence: 57%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Diversity of Melissococcus plutonius from Honeybee Larvae in Japan and Experimental Reproduction of European Foulbrood with Cultured Atypical Isolates

Arai

Tominaga²,

et al. 2012

PLoS ONE

154

View full text Add to dashboard Cite

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“…The bacterium is widely distributed ( Bailey, 1981 ) and can also infect and kill the brood of the Eastern honey bee ( Apis cerana ) and the Himalayan honey bee ( Apis laboriosa; Bailey, 1974 ; Allen et al, 1990 ). Although early reports suggested a highly genetically homogenous population ( Djordjevic et al , 1999 ), isolates of M. plutonius have been shown to vary in biochemical, serological and in vitro virulence characteristics ( Allen and Ball, 1993 ; Arai et al , 2012 ; Takamatsu et al , 2013 ).…”

Section: Introductionmentioning

confidence: 99%

Molecular epidemiology and population structure of the honey bee brood pathogenMelissococcus plutonius

et al. 2014

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Melissococcus plutonius is the causative agent of European foulbrood (EFB), which is a serious brood disease of the European honey bee (Apis mellifera). EFB remains a threat because of a poor understanding of disease epidemiology. We used a recently published multi-locus sequence typing method to characterise 206 M. plutonius isolates recovered from outbreaks in England and Wales over the course of 2 years. We detected 15 different sequence types (STs), which were resolved by eBURST and phylogenetic analysis into three clonal complexes (CCs) 3, 12 and 13. Single and double locus variants within CC3 were the most abundant and widespread genotypes, accounting for 85% of the cases. In contrast, CCs 12 and 13 were rarer and predominantly found in geographical regions of high sampling intensity, consistent with a more recent introduction and localised spread. K-function analysis and interpoint distance tests revealed significant geographical clustering in five common STs, but pointed to different dispersal patterns between STs. We noted that CCs appeared to vary in pathogenicity and that infection caused by the more pathogenic variants is more likely to lead to honey bee colony destruction, as opposed to treatment. The importance of these findings for improving our understanding of disease aetiology and control are discussed.

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“…This species had been thought to be remarkably homogeneous based on morphological, physiological, immunological and genetic studies [1, 7, 9]. However, Arai et al [2] recently reported the presence and prevalence of atypical M. plutonius , which are phenotypically and genetically distinguished from typical M. plutonius strains, in Japan.…”

mentioning

confidence: 99%

Development of Duplex PCR Assay for Detection and Differentiation of Typical and Atypical <i>Melissococcus plutonius</i> strains

Arai¹,

Miyoshi‐Akiyama

Okumura

et al. 2014

The Journal of Veterinary Medical Science

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Melissococcus plutonius is the causative agent of an important honeybee disease, European foulbrood (EFB). In addition to M. plutonius strains with typical characteristics (typical M. plutonius), we recently reported the presence of atypical M. plutonius, which are phenotypically and genetically distinguished from typical M. plutonius. Because typical and atypical M. plutonius may have different pathogenic mechanisms, differentiation of these two types is very important for diagnosis and more effective control of EFB. In this study, therefore, a duplex PCR assay was developed to detect and differentiate typical and atypical M. plutonius rapidly and easily. On the basis of the results of comparative genomic analyses, we selected Na+/H+ antiporter gene and Fur family transcriptional regulator gene as targets for detection of typical and atypical strains, respectively, by PCR. Under optimized conditions, the duplex PCR system using the designed primers successfully detected and differentiated all typical and atypical M. plutonius strain/isolates tested, while no product was generated from any other bacterial strains/isolates used in this study, including those isolated from healthy honeybee larval guts. Detection limits of the PCR were 50 copies of chromosome/reaction for both types, and it could detect typical and atypical M. plutonius directly from diseased honeybee larvae. Moreover, the duplex PCR diagnosed mixed infections with both M. plutonius types more precisely than standard culture methods. These results indicate that the duplex PCR assay developed in this study is extremely useful for precise diagnosis and epidemiological study of EFB.

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Geographically diverse Australian isolates ofMelissococcus plutonexhibit minimal genotypic diversity by restriction endonuclease analysis

Cited by 25 publications

References 13 publications

Diversity of Melissococcus plutonius from Honeybee Larvae in Japan and Experimental Reproduction of European Foulbrood with Cultured Atypical Isolates

Diversity of Melissococcus plutonius from Honeybee Larvae in Japan and Experimental Reproduction of European Foulbrood with Cultured Atypical Isolates

Molecular epidemiology and population structure of the honey bee brood pathogenMelissococcus plutonius

Development of Duplex PCR Assay for Detection and Differentiation of Typical and Atypical <i>Melissococcus plutonius</i> strains

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