PCR fingerprinting of whole genomes, the spacers between the 16S and 23S rRNA genes and of intergenic tRNA gene regions reveal a different intraspecific genomic variability of Bacillus cereus and Bacillus licheniformis

Daffonchio, D.; Borin, Sara; Frova, G.; Manachini, P. L.; Sorlini, C.

doi:10.1099/00207713-48-3-1081

Cited by 63 publications

(101 citation statements)

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“…However, we also determined that some physiological, morphological and biochemical characteristics of our isolate differ from those of Anoxybacillus flavithermus DSM 2641 T . In addition, Daffonchio et al (1998) showed that the 16S-23S ITS of Bacillus cereus are well conserved in terms of length; in contrast, bacilli such as Bacillus licheniformis and Bacillus subtilis have at least two different ITS fingerprints.…”

Section: Enzyme Assaysmentioning

confidence: 99%

Anoxybacillus gonensis sp. nov., a moderately thermophilic, xylose-utilizing, endospore-forming bacterium

Beldüz

Dülger

Demirbağ

2003

International Journal of Systematic and Evolutionary Microbiology

109

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Seven closely related xylanolytic, thermophilic bacilli were isolated from mud and water samples from the Gonen and Diyadin hot springs, respectively located in the Turkish provinces of Balikesir and Agri. On the basis of morphology and biochemical characteristics, one of the isolates, designated strain G2 T , was studied further. Strain G2 T is a xylanolytic, sporulating, Grampositive, rod-shaped bacterium. The isolate is a thermophilic (optimum temperature for growth, 55-60˚C), facultative anaerobe that grows on a wide range of carbon sources, including glucose, starch, xylose and mannitol. It expressed a high level of xylose isomerase activity on xylose and also on glucose. 16S rRNA gene sequence analysis showed that this isolate resembled Anoxybacillus flavithermus DSM 2641 T (>97 % similarity), but 16S-23S rDNA internally transcribed spacer polymorphism PCR showed variation between DSM 2641 T and isolate G2 T . However, it is also known that analysis of 16S rRNA gene sequences may be insufficient to distinguish between some species. In DNA-DNA hybridization, thermophilic isolate G2 T showed relatedness of 53?4 % to A. flavithermus and about 45?0 % to Anoxybacillus pushchinoensis, indicating that it is distinct at the species level. On the basis of the evidence presented, it is proposed that strain G2 T (=NCIMB 13933 T =NCCB 100040 T ) be designated as the type strain of Anoxybacillus gonensis sp. nov.It is now over a century since thermophiles were first reported (Miquel, 1888). Over the years, a number of sporeforming thermophiles have been reported, mainly in the genera Bacillus and Clostridium (Guagliardi et al., 1996).In this study, we isolated some thermophilic bacilli from the Gonen and Diyadin hot springs, respectively located in the Turkish provinces of Balikesir and Agri. On the basis of preliminary experiments, a representative strain appeared to differ from other thermophilic bacilli with respect to the utilization of xylose; it was therefore characterized further. Xylose isomerase is an intracellular enzyme that catalyses the conversion of D-xylose to D-xylulose. Its practical significance stems from its ability to isomerize D-glucose to D-fructose. Therefore, this enzyme is often referred to as glucose isomerase and is widely used in industry for the production of high-fructose corn syrup.The present paper describes the isolation, morphological, physiological and biochemical profiles and 16S rRNA sequence of this strain and the results of DNA-DNA hybridization with close relatives and proposes that it represents a novel species of the genus Anoxybacillus (Pikuta et al., 2000), Anoxybacillus gonensis sp. nov. Isolation of strainsSeven Gram-positive rods were isolated from mud and water samples from the Gonen and Diyadin hot springs. The water temperature of these hot springs is around 70 uC. After collection, mud and water samples were used immediately for enrichment in nutrient broth at 60-70 uC. Oneday-old enrichment cultures were repeatedly subcultured in 10 ml nutrient broth and streaked on ...

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Section: Enzyme Assaysmentioning

confidence: 99%

Anoxybacillus gonensis sp. nov., a moderately thermophilic, xylose-utilizing, endospore-forming bacterium

Beldüz

Dülger

Demirbağ

2003

International Journal of Systematic and Evolutionary Microbiology

109

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“…This simple profiling technique, known as ITS-PCR, allows characterizing complex microbial communities as a whole and offers a good phylogenetic resolution as, unlike the 16S and 23S genes, ITS undergoes a much milder selective pressure than its flanking genes do. Scarce evolutionary pressure upon spacers' sequences implies their sharp degree of polymorphism and hence high specificity, therefore making them suited to distinguish strains or ecotypes in the same species (Jensen et al 1993;Daffonchio et al 1998), whereas the comparison of 16S rDNA genes permits only to discriminate species belonging to the same genus (Weisburg et al 1991;Widmer et al 1998). Literature reports that ITS-PCR fingerprinting was successfully applied to the distinction of artisanal and industrial Mozzarella (Coppola et al 2001).…”

Section: Introductionmentioning

confidence: 99%

Genetic traceability of the geographical origin of typical Italian water buffalo Mozzarella cheese: a preliminary approach

et al. 2007

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Aims: To distinguish Italian Protected Designation of Origin (PDO) water buffalo Mozzarella from different producers on a molecular basis in relation to the place of manufacturing within the production district, and to develop a tool for genetic traceability of typical dairy products. Methods and Results: Microbial DNA was isolated from Mozzarella's governing liquid to amplify the whole microflora's ribosomal 16S–23S internal transcribed spacers (ITS)‐PCR fingerprinting by means of an original primer pair. Phylogenetic distance analyses were performed on the obtained electrophoretic band patterns by maximum parsimony and neighbour‐joining tree construction algorithms for discrete binary data, using a conventional bootstrap resampling test. The observed band profiles showed high repeatability and specificity, allowing unambiguous distinction of each sample; phylogenetic analyses yielded the same tree topology with good strength of nodal support. Moreover, a relationship between the genetic distances among samples and the actual geographical ones separating the respective producing dairies was observed. Conclusions: The genetic diversity of PDO water buffalo Mozzarella's microflora, observed by ITS‐PCR fingerprinting, can be exploited to discriminate cheeses from differently located dairies. Significance and Impact of the Study: Given the increasing importance of food traceability for safety, quality and typicalness issues, the ITS‐PCR fingerprinting protocol described here may represent a suitable tool for tracing the geographical origin of Italian Mozzarella.

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“…Therefore, some authors have emphasized the importance of tDNA-PCR as a tool for the identification of bacterial species. On the other hand, different profiles were detected by tDNA-PCR for S. aureus, Staphylococcus haemolyticus (Maes et al 1997), Bacillus licheniformis (Daffonchio et al 1998) and P. aeruginosa (Spacov et al 2006). In the present report tDNA-PCR allowed the discrimination of the clinical K. pneumoniae isolates from Brazil into three major patterns, with a predominance of the group that showed the same profile as the K. pneumoniae reference strains.…”

Section: Discussionmentioning

confidence: 68%

“…Its amplification pattern differed from related species analyzed, showing that this technique is efficient for the molecular identification of K. pneumoniae isolates. In other reports, PCR ribotyping also did not show polymorphism of amplification patterns for different isolates of Pseudomonas aeruginosa (Agodi et al 2000), Bacillus cereus (Daffonchio et al 1998) and for various species of the genus Salmonella (Lagatolla et al 1996), indicating that this method is efficient for the identification of these bacteria. On the other hand, isolates of Enterobacter cloacae (Clementino et al 2001), and Staphylococcus aureus (Pereira et al 2002), exhibit a high degree of polymorphism, and PCR ribotyping has therefore been indicated as a tool for the epidemiological study of these species.…”

Section: Discussionmentioning

confidence: 99%

Application of PCR ribotyping and tDNA-PCR for Klebsiella pneumoniae identification

Lópes

Rodrigues

Clementino

et al. 2007

Mem. Inst. Oswaldo Cruz

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PCR fingerprinting of whole genomes, the spacers between the 16S and 23S rRNA genes and of intergenic tRNA gene regions reveal a different intraspecific genomic variability of Bacillus cereus and Bacillus licheniformis

Cited by 63 publications

References 0 publications

Anoxybacillus gonensis sp. nov., a moderately thermophilic, xylose-utilizing, endospore-forming bacterium

Anoxybacillus gonensis sp. nov., a moderately thermophilic, xylose-utilizing, endospore-forming bacterium

Genetic traceability of the geographical origin of typical Italian water buffalo Mozzarella cheese: a preliminary approach

Application of PCR ribotyping and tDNA-PCR for Klebsiella pneumoniae identification

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