DNA restriction endonuclease profiles and typing of geographically diverse isolates of<i>Bacillus larvae</i>

Djordjevic, Steven P.; Ho-Shon, Michael; Hornitzky, M.

doi:10.1080/00218839.1994.11100856

Cited by 31 publications

(18 citation statements)

References 20 publications

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“…The two primary bacterial pathogens of honeybee colonies (M. pluton and P. larvae subsp. larvae) exhibit marked genetic, protein, and antigenic homogeneity, as determined by REA, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and immunoblot analyses (7,9,16), which suggests that these organisms evolved so that they form close host-parasite relationships with honeybee larvae. Although isolates 5, 9, 16, 24, and 26 through 28, which represent 23% of our isolates, were clonally related, biochemically they were among the most diverse isolates.…”

Section: Discussionmentioning

confidence: 99%

“…Total cellular DNA of 30 P. alvei isolates was extracted and purified by using the procedure developed for isolation of DNA from M. pluton and other gram-positive bacteria (7,8).…”

Section: Methodsmentioning

confidence: 99%

“…When restriction endonuclease analysis (REA) and immunoblot analysis were used, high levels of genetic and antigenic homogeneity were observed among geographically diverse Australian isolates of the primary honeybee pathogens P. larvae subsp. larvae (7,16) and M. pluton (9). There have been no reports of the levels of genetic heterogeneity among geographically diverse isolates of P. alvei.…”

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confidence: 99%

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Genetic and Biochemical Diversity among Isolates of Paenibacillus alvei Cultured from Australian Honeybee ( Apis mellifera ) Colonies

Djordjevic

Forbes

Smith

et al. 2000

Appl Environ Microbiol

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Twenty-five unique CfoI-generated whole-cell DNA profiles were identified in a study of 30 Paenibacillus alvei isolates cultured from honey and diseased larvae collected from honeybee (Apis mellifera) colonies in geographically diverse areas in Australia. The fingerprint patterns were highly variable and readily discernible from one another, which highlighted the potential of this method for tracing the movement of isolates in epidemiological studies. 16S rRNA gene fragments (length, 1,416 bp) for all 30 isolates were enzymatically amplified by PCR and subjected to restriction analysis with DraI, HinfI, CfoI, AluI, FokI, and RsaI. With each enzyme the restriction profiles of the 16S rRNA genes from all 30 isolates were identical (one restriction fragment length polymorphism [RFLP] was observed in the HinfI profile of the 16S rRNA gene from isolate 17), which confirmed that the isolates belonged to the same species. The restriction profiles generated by using DraI, FokI, and HinfI differentiated P. alvei from the phylogenetically closely related species Paenibacillus macerans and Paenibacillus macquariensis. Alveolysin gene fragments (length, 1,555 bp) were enzymatically amplified from some of the P. alvei isolates (19 of 30 isolates), and RFLP were detected by using the enzymes CfoI, Sau3AI, and RsaI. Extrachromosomal DNA ranging in size from 1 to 10 kb was detected in 17 of 30 (57%) P. alvei whole-cell DNA profiles. Extensive biochemical heterogeneity was observed among the 28 P. alvei isolates examined with the API 50CHB system. All of these isolates were catalase, oxidase, and VogesProskauer positive and nitrate negative, and all produced acid when glycerol, esculin, and maltose were added. The isolates produced variable results for 16 of the 49 biochemical tests; negative reactions were recorded in the remaining 30 assays. The genetic and biochemical heterogeneity in P. alvei isolates may be a reflection of adaptation to the special habitats in which they originated.Paenibacillus alvei is a spore-forming bacterium that swarms vigorously on routine culture media. P. alvei, Enterococcus faecalis, Enterococcus faecium, and Achromobacter eurydice are often recovered from diseased larvae obtained from honeybee (Apis mellifera) colonies infected with Melissococcus pluton, the causative agent of European foulbrood (EFB) (3). In Australia, P. alvei is the third-most common bacterium detected in honeybee colonies, and E. faecalis, E. faecium, and A. eurydice are rarely recovered from EFB-affected colonies (17, 18). P. alvei can produce signs in larvae that are similar to the signs produced by Paenibacillus larvae subsp. larvae, which causes American foulbrood, the other major bacterial disease of honeybees.Several studies based on comparative analyses of 16S rRNA gene sequences of different Bacillus species revealed five phylogenetically distinct clusters (groups 1 through 5), which confirmed that this genus is genetically heterogeneous and in need of extensive taxonomic revision (1, 27). Furthermore, the authors sug...

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Section: Discussionmentioning

confidence: 99%

“…Total cellular DNA of 30 P. alvei isolates was extracted and purified by using the procedure developed for isolation of DNA from M. pluton and other gram-positive bacteria (7,8).…”

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Genetic and Biochemical Diversity among Isolates of Paenibacillus alvei Cultured from Australian Honeybee ( Apis mellifera ) Colonies

Djordjevic

Forbes

Smith

et al. 2000

Appl Environ Microbiol

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“…In two recent studies, restriction fragment length polymorphism (RFLP) analysis was used to differentiate between different isolates of P. l. larvae (Djordjevic et al, 1994) or Paenibacillus alvei (P. alvei) (Djordjevic et al, 2000). CfoI-generated whole-cell DNA profiles that showed a very high degree of heterogeneity for both P. alvei and P. l. larvae, made it difficult to convincingly define clonal isolates.…”

Section: Introductionmentioning

confidence: 99%

The use of repetitive element PCR fingerprinting (rep-PCR) for genetic subtyping of German field isolates of Paenibacillus larvae subsp. larvae

Genersch

Otten²

2003

Apidologie

145

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-Studies using the repetitive element PCR fingerprinting technique (rep-PCR) revealed that BOX A1R-, MBO REP1-and ERIC-primers reproducibly generate distinctive DNA fingerprints from Paenibacillus larvae subsp. larvae. Four different genetic subtypes of P. larvae were identified in Germany using the three primers. Correlating the results from genetic and morphological subtyping, the flat and transparent morphotype could be assigned to one genetic group. Geographic evaluation of our results demonstrated that the different genetic subtypes appeared in clusters correlating with different outbreaks of American foulbrood. Taken together, our results indicated that rep-PCR performed with a combination of BOX A1R-and MBO REP1-primers will be an effective tool for establishing a molecular epidemiology of P. l. larvae.American foulbrood / Paenibacillus larvae larvae / DNA-fingerprinting / genetic subtyping / molecular epidemiology

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“…Acrylamide solutions were polymerised. with an excess of ammonium persulphate (1.25 mg ml-l) (Sigma) and TEMED (1 p1 ml-l) (Bio-Rad) as described by Djordjevic et al (1994). Molecular weight markers used were a combination of DNA fragments of known size generated by the cleavage of Bacillus subtilus phage SPP-1 DNA (Progen) with EcoRI, lambda phage DNA (Boehringer) digested wi.th HindIII, plasmid pbluescribe DNA (Stratagene) digested with PstI and bromophenol blue which migrates at approximately 100 bp in 3.5'Y1 polyacrylar-nide gels.…”

Section: Methodsmentioning

confidence: 99%

Restriction endonuclease analysis of atypical Aeromonas salmonicida isolates from goldfish Carassius auratus, silver perch Bidyanus bidyanus, and greenback flounder Rhombosolea tapirina in Australia

Whittington¹,

Djordjevic²,

Carson³

et al. 1995

Dis. Aquat. Org.

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Total genomic DNA samples of atypical isolates of Aeromonas salmonicida from goldfish Carassius auratus L., silver perch Bidyanus bidyanus (Mitchell), and greenback flounder Rhombosolea tapirina Giinther from Australia and isolates from goldfish from the USA and Singapore were digested with the restriction enzyme Cfol, separated In 3.5% acrylamide gels and visualised by silver staining. Isolates from goldfish and silver perch comprised an homogenous group while those from greenback flounder formed a second group. Within the goldfish and silver perch isolates there were minor differences in restriction endonuclease fragment pattern. Isolates from Australian goldfish obtained between 1977 and 1989 were indistinguishable from isolates from goldfish from the USA but slightly different from an isolate from a Singaporean goldfish. These and other data suggest that A. salmonicidaprobably crossed a species barrier from goldfish to silver perch on the same farm but that farmed greenback flounder In Tasmania have acquired a distinct straln of A. saln~onicida probably from wild-caught marine fish.

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DNA restriction endonuclease profiles and typing of geographically diverse isolates ofBacillus larvae

Cited by 31 publications

References 20 publications

Genetic and Biochemical Diversity among Isolates of Paenibacillus alvei Cultured from Australian Honeybee ( Apis mellifera ) Colonies

Genetic and Biochemical Diversity among Isolates of Paenibacillus alvei Cultured from Australian Honeybee ( Apis mellifera ) Colonies

The use of repetitive element PCR fingerprinting (rep-PCR) for genetic subtyping of German field isolates of Paenibacillus larvae subsp. larvae

Restriction endonuclease analysis of atypical Aeromonas salmonicida isolates from goldfish Carassius auratus, silver perch Bidyanus bidyanus, and greenback flounder Rhombosolea tapirina in Australia

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