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MSP016 The genetic determinant of intrinsic quinolone resistance in Fusobacterium nucleatum ssp. canifelium
Wissenschaftliches Programm 55. DGHM-Tagung 29. September-l. Oktober 2003 in Dresden Abstracts - Poster
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Salmonella enterica, serotype Enteritidis,
phagetype 4: a comparative analysis of genomic DNA fragment pattern (PFGE) and ribotyping for epidemiological subdifferentiation Prager, R}; Gericke, B.~; Rabsch, W.1; Tsch~pe, H }
~Robert Koch Institute; Wernigerode Branch S. enterica Serovar Enteritidis (SE) PT 4 is worldwide the dominant epidemic strain among cases of salmonellosis. In Germany approximately 75 % of isolated strains of S. enteritidis (SE) belong to this phage type. Therefore, reliable and extensive discriminative methods are required in order to differentiate SE, PT4 strains. In particular, ribotyping has been found to be a suitable subdifferentiation of SE (PT4). The aim of this study was to evaluate the genetic heterogeneity of SE (PT4) using ribotyping in comparison with PFGE. A collection of 170 outbreak and non-outbreak strains of SE, PT4 isolated from humans, poultry, egg and poultry meat in Germany between 1986 and 2002 was selected and genetically fingerprinted in order to assess the use of these techniques for epidemiological investigations. PstT/SphI-ribotyping differentiated SE, PT4 into 18 different ribotypes. XbaI-PFGE showed 6, SpeI-PFGE showed 7 different macrorestriction profiles. In summary, it has been found that PstI/SphIribotyping is a more sensitive technique for identifying polymorphism within SE (PT4) than PFGE. The results open the possibility for identification of sources of infection and routes of transmission. Therefore, PstI/SphI-ribotyping of SE, PT4 should be an important component of surveillance programmes.
The genetic determinant of intrinsic quinolone resistance in Fusobacterium nucleatum ssp. canifefium Conrads, G.1; Citron, D ] ; Jang, S.3; Goldstein, E ] 1Universit~tsklinikum (RWTH-Aachen); Lehr- und Forschungsgebiet Orale Mikrobiologie und Immunologie 2UCLA; RM Alden Research Laboratory 3University of California at Davis; Veterinary Medical Teaching Hospital Fourteen strains of Gram-negative, anaerobic, fluoroquinolone-resistant, non-sporulating rods that were isolated from the normal oral cavity of cats and dogs, as well as from wounds in humans after cat- or dog-bites, and which phenotypically resembled Fusobacterium nucleaturn, were characterized by sequencing of the 16S-23S rDNA internal transcribed spacer (ITS) regions and 16S rDNA, phylogenetic analysis, and phenotypic tests. The results placed the novel strains as a distinct genotype or subspecies of F. nucleatum. The subspecies F. nucleatum subsp. canifelium subsp, nov. has been proposed, with strain ATCC BAA 689 T (DSM 15542T), a dog isolate, as the type strain. All strains of this new subspecies are intrinsically resistant (MIC >4g/mL) to levofloxacin, moxifloxacin,
http://www.dghm.org
326
Wissenschaftliches Programm 55. DGHM-Tagung 29. September-1. Oktober 2003 in Dresden
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Abstracts - Poster
gatifloxacin, gemifloxacin, and other fluoroquinolones. As carbonyl cyanide chlorophenylhydrazone (201Jg/ml) /CCCP), an efflux inhibitor, did not reduce the MICs, we amplified and sequenced the quinolone resistancedetermining region (QRDR) of gyrA in ATCC BAA 689 T and in the cat isolate ATCC BAA 690 (DSM 15543). In comparison to QRDR of the fluoroquinolone-susceptible subspecies nucleatum and vincentii, we found three single nucleotide mutations in both strains of the resistant subspecies canifelium. Most important for the level of resistance is Ser79 (polar amino acid, equivalent to E.coli Ser83 and B. fragilis Ser82) which was replaced by leucine, a hydrophobic and aliphatic amino acid. In addition, equivalent to Asp87 (in quinolone-susceptible E.coli strains), we found the hydrophobic glycine in the subspecies nucleatum and vincentii as well as the polar and charged arginine in subspecies canifelium. This report represents the first description of fluoroquinolone-resistance in F. nucleatum.
Emergence of antibiotic resistance among S. pneumoniae and S. pyogenes strains isolated from the respiratory tract infections of children in Germany, 2 0 0 2 Reinert, R.R.1; Neuberger, N.1; Cil, M.Y.1; Cremer, C}; Lemperle, M.1; Luetticken, R.Z; AI-Lahham, A. ~ ZNational Reference Center for Streptococci-AC; Medical Microbiology
Respiratory tract infections caused by S. pneumoniae and S. pyogenes are serious health problems worldwide. In a multicentre study including 10 clinical laboratories, a total of 241 S. pneumoniae and 236 S. pyogenes strains were collected in 2002. MICs were determined according to NCCLS by microbroth dilution method; macrolide resistance phenotypes of both pathogens were determined by double disk diffusion test. PCR of macrolide resistant determinants was performed according to standard methods. Resistance rates of S. pyogenes isolates were as follows (intermediate and resistant): Penicillin 0%, cefotaxime 0%, amoxicillin 0%, erythromycin A 14%, clindamycin 0%, gatifloxacin 0% and telithromycin 0%. S. pneumoniae isolates showed the following resistance rates (intermediate and resistant): Penicillin 5%, cefotaxime 1.7%, amoxicillin 0.8%, erythromycin 19.9%, clindamycin 7.5% and gatifloxacin 0%. The new ketolide telithromycin was 100% active against all S. pneumoniae (MIC 50, 0.016 mg/L; MIC 90 = 0.125 mg/L) and S. pyogenes (MIC 5o , 0.03 mg/L; MIC 9o, 0.5 rag/L). Of 48 macrolide-resistant pneumococcal strains 30 (62.5%) were M phenotypes possessing the mef(A) resistant determinants and 18 (37.5%) were cMLSB possessing the errn(B) resistance determinants. 33 isolates of S. pyogenes were macrolide resistant, among which 31 (93.9%) were M phenotypes. In conclusion, macrolide resistance among children is an increasing problem in Germany. Telithromycin was highly active against all S. pneumoniae and S. pyogenes isolates.
http://www.dghm.org
327
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Salmonella enterica, serotype Enteritidis,
phagetype 4: a comparative analysis of genomic DNA fragment pattern (PFGE) and ribotyping for epidemiological subdifferentiation Prager, R}; Gericke, B.~; Rabsch, W.1; Tsch~pe, H }
~Robert Koch Institute; Wernigerode Branch S. enterica Serovar Enteritidis (SE) PT 4 is worldwide the dominant epidemic strain among cases of salmonellosis. In Germany approximately 75 % of isolated strains of S. enteritidis (SE) belong to this phage type. Therefore, reliable and extensive discriminative methods are required in order to differentiate SE, PT4 strains. In particular, ribotyping has been found to be a suitable subdifferentiation of SE (PT4). The aim of this study was to evaluate the genetic heterogeneity of SE (PT4) using ribotyping in comparison with PFGE. A collection of 170 outbreak and non-outbreak strains of SE, PT4 isolated from humans, poultry, egg and poultry meat in Germany between 1986 and 2002 was selected and genetically fingerprinted in order to assess the use of these techniques for epidemiological investigations. PstT/SphI-ribotyping differentiated SE, PT4 into 18 different ribotypes. XbaI-PFGE showed 6, SpeI-PFGE showed 7 different macrorestriction profiles. In summary, it has been found that PstI/SphIribotyping is a more sensitive technique for identifying polymorphism within SE (PT4) than PFGE. The results open the possibility for identification of sources of infection and routes of transmission. Therefore, PstI/SphI-ribotyping of SE, PT4 should be an important component of surveillance programmes.
The genetic determinant of intrinsic quinolone resistance in Fusobacterium nucleatum ssp. canifefium Conrads, G.1; Citron, D ] ; Jang, S.3; Goldstein, E ] 1Universit~tsklinikum (RWTH-Aachen); Lehr- und Forschungsgebiet Orale Mikrobiologie und Immunologie 2UCLA; RM Alden Research Laboratory 3University of California at Davis; Veterinary Medical Teaching Hospital Fourteen strains of Gram-negative, anaerobic, fluoroquinolone-resistant, non-sporulating rods that were isolated from the normal oral cavity of cats and dogs, as well as from wounds in humans after cat- or dog-bites, and which phenotypically resembled Fusobacterium nucleaturn, were characterized by sequencing of the 16S-23S rDNA internal transcribed spacer (ITS) regions and 16S rDNA, phylogenetic analysis, and phenotypic tests. The results placed the novel strains as a distinct genotype or subspecies of F. nucleatum. The subspecies F. nucleatum subsp. canifelium subsp, nov. has been proposed, with strain ATCC BAA 689 T (DSM 15542T), a dog isolate, as the type strain. All strains of this new subspecies are intrinsically resistant (MIC >4g/mL) to levofloxacin, moxifloxacin,
http://www.dghm.org
326
Wissenschaftliches Programm 55. DGHM-Tagung 29. September-1. Oktober 2003 in Dresden
L
Abstracts - Poster
gatifloxacin, gemifloxacin, and other fluoroquinolones. As carbonyl cyanide chlorophenylhydrazone (201Jg/ml) /CCCP), an efflux inhibitor, did not reduce the MICs, we amplified and sequenced the quinolone resistancedetermining region (QRDR) of gyrA in ATCC BAA 689 T and in the cat isolate ATCC BAA 690 (DSM 15543). In comparison to QRDR of the fluoroquinolone-susceptible subspecies nucleatum and vincentii, we found three single nucleotide mutations in both strains of the resistant subspecies canifelium. Most important for the level of resistance is Ser79 (polar amino acid, equivalent to E.coli Ser83 and B. fragilis Ser82) which was replaced by leucine, a hydrophobic and aliphatic amino acid. In addition, equivalent to Asp87 (in quinolone-susceptible E.coli strains), we found the hydrophobic glycine in the subspecies nucleatum and vincentii as well as the polar and charged arginine in subspecies canifelium. This report represents the first description of fluoroquinolone-resistance in F. nucleatum.
Emergence of antibiotic resistance among S. pneumoniae and S. pyogenes strains isolated from the respiratory tract infections of children in Germany, 2 0 0 2 Reinert, R.R.1; Neuberger, N.1; Cil, M.Y.1; Cremer, C}; Lemperle, M.1; Luetticken, R.Z; AI-Lahham, A. ~ ZNational Reference Center for Streptococci-AC; Medical Microbiology
Respiratory tract infections caused by S. pneumoniae and S. pyogenes are serious health problems worldwide. In a multicentre study including 10 clinical laboratories, a total of 241 S. pneumoniae and 236 S. pyogenes strains were collected in 2002. MICs were determined according to NCCLS by microbroth dilution method; macrolide resistance phenotypes of both pathogens were determined by double disk diffusion test. PCR of macrolide resistant determinants was performed according to standard methods. Resistance rates of S. pyogenes isolates were as follows (intermediate and resistant): Penicillin 0%, cefotaxime 0%, amoxicillin 0%, erythromycin A 14%, clindamycin 0%, gatifloxacin 0% and telithromycin 0%. S. pneumoniae isolates showed the following resistance rates (intermediate and resistant): Penicillin 5%, cefotaxime 1.7%, amoxicillin 0.8%, erythromycin 19.9%, clindamycin 7.5% and gatifloxacin 0%. The new ketolide telithromycin was 100% active against all S. pneumoniae (MIC 50, 0.016 mg/L; MIC 90 = 0.125 mg/L) and S. pyogenes (MIC 5o , 0.03 mg/L; MIC 9o, 0.5 rag/L). Of 48 macrolide-resistant pneumococcal strains 30 (62.5%) were M phenotypes possessing the mef(A) resistant determinants and 18 (37.5%) were cMLSB possessing the errn(B) resistance determinants. 33 isolates of S. pyogenes were macrolide resistant, among which 31 (93.9%) were M phenotypes. In conclusion, macrolide resistance among children is an increasing problem in Germany. Telithromycin was highly active against all S. pneumoniae and S. pyogenes isolates.
http://www.dghm.org
327