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Maria Cruz Martin - One of the best experts on this subject based on the ideXlab platform.
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cytotoxin and pyrogenic toxin superantigen gene profiles of staphylococcus aureus associated with subclinical mastitis in dairy cows and relationships with macrorestriction genomic profiles
Journal of Clinical Microbiology, 2005Co-Authors: J M Fueyo, M R Rodicio, M C Mendoza, J Muniz, Miguel A Alvarez, Maria Cruz MartinAbstract:A set of 84 Staphylococcus aureus isolates collected from the milk of cows with subclinical mastitis in Asturias (a cattle region of Spain) and six control strains were tested for sequences of genes encoding hemolysins (hla, hlb, hld, hlg, and hlg-2), leukotoxins (lukPV, lukM, and lukED), toxic shock syndrome toxin (tst), and enterotoxins (sea to see, seg to ser, and seu) by conventional and multiplex PCR. It was found that 84, 83, 11, and 39 isolates carried some type of hl, luk, tst, or se gene, respectively, which were arranged in 14 exotoxin genotypes. All of the isolates were negative for lukPV, hlg, sea, sed, see, sej, sek, sep, seq, and ser. Two gene groupings could be related with pathogenicity islands—[lukED, seg, sei, sem, sen, seo ± seu] with Saβ-1 and [tst, sec, sel] with SaPIbov, present in 45 and 13.1% of the isolates, respectively—while 11.9% of them carried both islands. Only one contained seb (together with υSaβ-1), and another contained seh (together with lukED). The isolates were also analyzed by pulsed-field gel electrophoresis performed with SmaI. Thirty-nine SmaI profiles (similarity coefficient [S] = 0.94 to 0.21) were differentiated; 12, 1, and 10 of these, respectively, were generated by isolates presumptively carrying Saβ-1, SaPIbov, or both. Five SmaI profiles (S ≥ 0.8) formed a cluster, which contained 20 and 10 isolates carrying one (υSaβ-1) or both islands. These data show the high frequency of genes encoding cytotoxins and pyrogenic toxin superantigens, their relationship with pathogenicity islands, and their distribution among a diversity of genetic types of S. aureus related to subclinical mastitis.
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cytotoxin and pyrogenic toxin superantigen gene profiles of staphylococcus aureus associated with subclinical mastitis in dairy cows and relationships with macrorestriction genomic profiles
Journal of Clinical Microbiology, 2005Co-Authors: J M Fueyo, M R Rodicio, M C Mendoza, J Muniz, Miguel A Alvarez, Maria Cruz MartinAbstract:A set of 84 Staphylococcus aureus isolates collected from the milk of cows with subclinical mastitis in Asturias (a cattle region of Spain) and six control strains were tested for sequences of genes encoding hemolysins (hla, hlb, hld, hlg, and hlg-2), leukotoxins (lukPV, lukM, and lukED), toxic shock syndrome toxin (tst), and enterotoxins (sea to see, seg to ser, and seu) by conventional and multiplex PCR. It was found that 84, 83, 11, and 39 isolates carried some type of hl, luk, tst, or se gene, respectively, which were arranged in 14 exotoxin genotypes. All of the isolates were negative for lukPV, hlg, sea, sed, see, sej, sek, sep, seq, and ser. Two gene groupings could be related with pathogenicity islands—[lukED, seg, sei, sem, sen, seo ± seu] with Saβ-1 and [tst, sec, sel] with SaPIbov, present in 45 and 13.1% of the isolates, respectively—while 11.9% of them carried both islands. Only one contained seb (together with υSaβ-1), and another contained seh (together with lukED). The isolates were also analyzed by pulsed-field gel electrophoresis performed with SmaI. Thirty-nine SmaI profiles (similarity coefficient [S] = 0.94 to 0.21) were differentiated; 12, 1, and 10 of these, respectively, were generated by isolates presumptively carrying Saβ-1, SaPIbov, or both. Five SmaI profiles (S ≥ 0.8) formed a cluster, which contained 20 and 10 isolates carrying one (υSaβ-1) or both islands. These data show the high frequency of genes encoding cytotoxins and pyrogenic toxin superantigens, their relationship with pathogenicity islands, and their distribution among a diversity of genetic types of S. aureus related to subclinical mastitis.
J M Fueyo - One of the best experts on this subject based on the ideXlab platform.
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cytotoxin and pyrogenic toxin superantigen gene profiles of staphylococcus aureus associated with subclinical mastitis in dairy cows and relationships with macrorestriction genomic profiles
Journal of Clinical Microbiology, 2005Co-Authors: J M Fueyo, M R Rodicio, M C Mendoza, J Muniz, Miguel A Alvarez, Maria Cruz MartinAbstract:A set of 84 Staphylococcus aureus isolates collected from the milk of cows with subclinical mastitis in Asturias (a cattle region of Spain) and six control strains were tested for sequences of genes encoding hemolysins (hla, hlb, hld, hlg, and hlg-2), leukotoxins (lukPV, lukM, and lukED), toxic shock syndrome toxin (tst), and enterotoxins (sea to see, seg to ser, and seu) by conventional and multiplex PCR. It was found that 84, 83, 11, and 39 isolates carried some type of hl, luk, tst, or se gene, respectively, which were arranged in 14 exotoxin genotypes. All of the isolates were negative for lukPV, hlg, sea, sed, see, sej, sek, sep, seq, and ser. Two gene groupings could be related with pathogenicity islands—[lukED, seg, sei, sem, sen, seo ± seu] with Saβ-1 and [tst, sec, sel] with SaPIbov, present in 45 and 13.1% of the isolates, respectively—while 11.9% of them carried both islands. Only one contained seb (together with υSaβ-1), and another contained seh (together with lukED). The isolates were also analyzed by pulsed-field gel electrophoresis performed with SmaI. Thirty-nine SmaI profiles (similarity coefficient [S] = 0.94 to 0.21) were differentiated; 12, 1, and 10 of these, respectively, were generated by isolates presumptively carrying Saβ-1, SaPIbov, or both. Five SmaI profiles (S ≥ 0.8) formed a cluster, which contained 20 and 10 isolates carrying one (υSaβ-1) or both islands. These data show the high frequency of genes encoding cytotoxins and pyrogenic toxin superantigens, their relationship with pathogenicity islands, and their distribution among a diversity of genetic types of S. aureus related to subclinical mastitis.
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cytotoxin and pyrogenic toxin superantigen gene profiles of staphylococcus aureus associated with subclinical mastitis in dairy cows and relationships with macrorestriction genomic profiles
Journal of Clinical Microbiology, 2005Co-Authors: J M Fueyo, M R Rodicio, M C Mendoza, J Muniz, Miguel A Alvarez, Maria Cruz MartinAbstract:A set of 84 Staphylococcus aureus isolates collected from the milk of cows with subclinical mastitis in Asturias (a cattle region of Spain) and six control strains were tested for sequences of genes encoding hemolysins (hla, hlb, hld, hlg, and hlg-2), leukotoxins (lukPV, lukM, and lukED), toxic shock syndrome toxin (tst), and enterotoxins (sea to see, seg to ser, and seu) by conventional and multiplex PCR. It was found that 84, 83, 11, and 39 isolates carried some type of hl, luk, tst, or se gene, respectively, which were arranged in 14 exotoxin genotypes. All of the isolates were negative for lukPV, hlg, sea, sed, see, sej, sek, sep, seq, and ser. Two gene groupings could be related with pathogenicity islands—[lukED, seg, sei, sem, sen, seo ± seu] with Saβ-1 and [tst, sec, sel] with SaPIbov, present in 45 and 13.1% of the isolates, respectively—while 11.9% of them carried both islands. Only one contained seb (together with υSaβ-1), and another contained seh (together with lukED). The isolates were also analyzed by pulsed-field gel electrophoresis performed with SmaI. Thirty-nine SmaI profiles (similarity coefficient [S] = 0.94 to 0.21) were differentiated; 12, 1, and 10 of these, respectively, were generated by isolates presumptively carrying Saβ-1, SaPIbov, or both. Five SmaI profiles (S ≥ 0.8) formed a cluster, which contained 20 and 10 isolates carrying one (υSaβ-1) or both islands. These data show the high frequency of genes encoding cytotoxins and pyrogenic toxin superantigens, their relationship with pathogenicity islands, and their distribution among a diversity of genetic types of S. aureus related to subclinical mastitis.
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enterotoxins and toxic shock syndrome toxin in staphylococcus aureus recovered from human nasal carriers and manually handled foods epidemiological and genetic findings
Microbes and Infection, 2005Co-Authors: J M Fueyo, Carmen M Mendoza, Cruz M MartinAbstract:A set of 269 Staphylococcus aureus isolates recovered from nasal carriers and manually handled foods in a region of Spain was analyzed for pyrogenic toxin production and toxin genes. Fifty-seven isolates producing at least one of four enterotoxins (SEA, SEB, SEC, SED), 10 isolates producing only toxic shock syndrome toxin (TSST-1), and 10 isolates producing both toxin types were found. The 77 toxigenic isolates were discriminated into 36 SmaI genomic and 13 EcoRI plasmid profiles. A strong relationship between toxin profiles with both SmaI genomic and EcoRI plasmid profiles was revealed. SmaI genomic profiles showing six or less mismatching fragments and similarity coefficient > or =0.7 were included in a lineage. Eight lineages were differentiated; six of them grouped both human and food isolates and two of these also included outbreak-implicated isolates. Two lineages, represented by TSST-SEA and TSST-1, on the one hand, and SEC and SEC-SED isolates, on the other hand, were the most frequent, but only the second was outbreak-related. When SmaI genomic and EcoRI plasmid profiles were hybridized with tst, sea, seb, and sec toxin probes, it was observed that each probe mapped on a different SmaI fragment from isolates falling into the same lineage. All of the probes only mapped on genomic fragments, but sed also mapped on three plasmid fragments. When sej and ser probes were included, they mapped together with sed on the chromosome and on the plasmids. Two plasmids (ca. 33 and 36 kb) carried the expected sed-sej-ser genes, while the other (ca. 53.5 kb) carried sed-sej and ser-like genes. The latter plasmid and the chromosomal location of sed-sej-ser are new findings from this study.
M R Rodicio - One of the best experts on this subject based on the ideXlab platform.
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high heterogeneity within methicillin resistant staphylococcus aureus st398 isolates defined by cfr9i macrorestriction pulsed field gel electrophoresis profiles and spa and sccmec types
Applied and Environmental Microbiology, 2010Co-Authors: M A Argudin, A Fetsch, B A Tenhagen, Jens A Hammerl, Stefan Hertwig, J Kowall, M R Rodicio, Annemarie Kasbohrer, Reiner HelmuthAbstract:During recent years, the animal-associated methicillin-resistant Staphylococcus aureus clone ST398 has extensively been studied. The DNA of these isolates turned out to be refractory to SmaI restriction, and consequently, SmaI is unsuitable for subtyping this clone by standard pulsed-field gel electrophoresis (PFGE). Very recently, ST398 DNA was shown to be digested by Cfr9I, a neoschizomer of SmaI. In the present study, we employed Cfr9I PFGE on 100 German and 5 Dutch ST398 isolates and compared their PFGE profiles, protein A gene variable repeat regions (spa types), and types of the staphylococcal cassette chromosome mec (SCCmec). The isolates (from healthy carrier pigs, clinical samples from pigs, dust from farms, milk, and meat) were assigned to 35 profiles, which were correlated to the SCCmec type. A dendrogram with the Cfr9I patterns assigned all profiles to two clusters. Cluster A grouped nearly all isolates with SCCmec type V, and cluster B comprised all SCCmec type IVa and V* (a type V variant first identified as III) carriers plus one isolate with SCCmec type V. Both clusters also grouped methicillin-susceptible S. aureus isolates. The association of the majority of isolates with SCCmec type V in one large cluster indicated the presence of a successful subclone within the clonal complex CC398 from pigs, which has diversified. In general, the combination of Cfr9I PFGE with spa and SCCmec typing demonstrated the heterogeneity of the series analyzed and can be further used for outbreak investigations and traceability studies of the MRSA ST398 emerging clone.
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the emerging methicillin resistant staphylococcus aureus st398 clone can easily be typed using the cfr9i SmaI neoschizomer
Letters in Applied Microbiology, 2010Co-Authors: M A Argudin, M R Rodicio, Beatriz GuerraAbstract:Aim: To establish a PFGE protocol using Cfr9I, neoschizomer of SmaI, for typing of Staphylococcus aureus isolates belonging to the emerging MRSA ST398 clone. Methods and Results: Staphylococcus aureus ST398 and non-ST398 isolates were analysed using the PFGE conditions recommended by the HARMONY consensus protocol. Genomic DNA of non-ST398 isolates could be digested with SmaI, XmaI (also a SmaI-neoschizomer) and Cfr9I. The DNA of SmaI-nontypeable ST398 isolates was partially resistant to XmaI, but could be digested with Cfr9I. By PCR-amplification/sequencing, the presence of a novel C5-cytosine methyltransferase gene (sauST398M) was detected in the ST398 isolates. The encoded enzyme, which shows high similarity with C5-cytosine methyltransferases that modify the CCCGGG recognition sequence, could be responsible for the different restriction results. Conclusion: SmaI-PFGE is regarded as the ‘gold standard’ for typing S. aureus. Because of different susceptibility of the GGGCCC recognition sites of the ST398 DNA against SmaI, XmaI and Cfr9I, the proposed protocol is a valuable tool for ST398 typing. Significance and Impact of the Study: The use of this protocol allows the comparison of results from SmaI-nontypeable isolates with S. aureus SmaI-PFGE databases and can be applied for outbreak investigations and traceability studies of this emerging MRSA clone.
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cytotoxin and pyrogenic toxin superantigen gene profiles of staphylococcus aureus associated with subclinical mastitis in dairy cows and relationships with macrorestriction genomic profiles
Journal of Clinical Microbiology, 2005Co-Authors: J M Fueyo, M R Rodicio, M C Mendoza, J Muniz, Miguel A Alvarez, Maria Cruz MartinAbstract:A set of 84 Staphylococcus aureus isolates collected from the milk of cows with subclinical mastitis in Asturias (a cattle region of Spain) and six control strains were tested for sequences of genes encoding hemolysins (hla, hlb, hld, hlg, and hlg-2), leukotoxins (lukPV, lukM, and lukED), toxic shock syndrome toxin (tst), and enterotoxins (sea to see, seg to ser, and seu) by conventional and multiplex PCR. It was found that 84, 83, 11, and 39 isolates carried some type of hl, luk, tst, or se gene, respectively, which were arranged in 14 exotoxin genotypes. All of the isolates were negative for lukPV, hlg, sea, sed, see, sej, sek, sep, seq, and ser. Two gene groupings could be related with pathogenicity islands—[lukED, seg, sei, sem, sen, seo ± seu] with Saβ-1 and [tst, sec, sel] with SaPIbov, present in 45 and 13.1% of the isolates, respectively—while 11.9% of them carried both islands. Only one contained seb (together with υSaβ-1), and another contained seh (together with lukED). The isolates were also analyzed by pulsed-field gel electrophoresis performed with SmaI. Thirty-nine SmaI profiles (similarity coefficient [S] = 0.94 to 0.21) were differentiated; 12, 1, and 10 of these, respectively, were generated by isolates presumptively carrying Saβ-1, SaPIbov, or both. Five SmaI profiles (S ≥ 0.8) formed a cluster, which contained 20 and 10 isolates carrying one (υSaβ-1) or both islands. These data show the high frequency of genes encoding cytotoxins and pyrogenic toxin superantigens, their relationship with pathogenicity islands, and their distribution among a diversity of genetic types of S. aureus related to subclinical mastitis.
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cytotoxin and pyrogenic toxin superantigen gene profiles of staphylococcus aureus associated with subclinical mastitis in dairy cows and relationships with macrorestriction genomic profiles
Journal of Clinical Microbiology, 2005Co-Authors: J M Fueyo, M R Rodicio, M C Mendoza, J Muniz, Miguel A Alvarez, Maria Cruz MartinAbstract:A set of 84 Staphylococcus aureus isolates collected from the milk of cows with subclinical mastitis in Asturias (a cattle region of Spain) and six control strains were tested for sequences of genes encoding hemolysins (hla, hlb, hld, hlg, and hlg-2), leukotoxins (lukPV, lukM, and lukED), toxic shock syndrome toxin (tst), and enterotoxins (sea to see, seg to ser, and seu) by conventional and multiplex PCR. It was found that 84, 83, 11, and 39 isolates carried some type of hl, luk, tst, or se gene, respectively, which were arranged in 14 exotoxin genotypes. All of the isolates were negative for lukPV, hlg, sea, sed, see, sej, sek, sep, seq, and ser. Two gene groupings could be related with pathogenicity islands—[lukED, seg, sei, sem, sen, seo ± seu] with Saβ-1 and [tst, sec, sel] with SaPIbov, present in 45 and 13.1% of the isolates, respectively—while 11.9% of them carried both islands. Only one contained seb (together with υSaβ-1), and another contained seh (together with lukED). The isolates were also analyzed by pulsed-field gel electrophoresis performed with SmaI. Thirty-nine SmaI profiles (similarity coefficient [S] = 0.94 to 0.21) were differentiated; 12, 1, and 10 of these, respectively, were generated by isolates presumptively carrying Saβ-1, SaPIbov, or both. Five SmaI profiles (S ≥ 0.8) formed a cluster, which contained 20 and 10 isolates carrying one (υSaβ-1) or both islands. These data show the high frequency of genes encoding cytotoxins and pyrogenic toxin superantigens, their relationship with pathogenicity islands, and their distribution among a diversity of genetic types of S. aureus related to subclinical mastitis.
M A Argudin - One of the best experts on this subject based on the ideXlab platform.
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high heterogeneity within methicillin resistant staphylococcus aureus st398 isolates defined by cfr9i macrorestriction pulsed field gel electrophoresis profiles and spa and sccmec types
Applied and Environmental Microbiology, 2010Co-Authors: M A Argudin, A Fetsch, B A Tenhagen, Jens A Hammerl, Stefan Hertwig, J Kowall, M R Rodicio, Annemarie Kasbohrer, Reiner HelmuthAbstract:During recent years, the animal-associated methicillin-resistant Staphylococcus aureus clone ST398 has extensively been studied. The DNA of these isolates turned out to be refractory to SmaI restriction, and consequently, SmaI is unsuitable for subtyping this clone by standard pulsed-field gel electrophoresis (PFGE). Very recently, ST398 DNA was shown to be digested by Cfr9I, a neoschizomer of SmaI. In the present study, we employed Cfr9I PFGE on 100 German and 5 Dutch ST398 isolates and compared their PFGE profiles, protein A gene variable repeat regions (spa types), and types of the staphylococcal cassette chromosome mec (SCCmec). The isolates (from healthy carrier pigs, clinical samples from pigs, dust from farms, milk, and meat) were assigned to 35 profiles, which were correlated to the SCCmec type. A dendrogram with the Cfr9I patterns assigned all profiles to two clusters. Cluster A grouped nearly all isolates with SCCmec type V, and cluster B comprised all SCCmec type IVa and V* (a type V variant first identified as III) carriers plus one isolate with SCCmec type V. Both clusters also grouped methicillin-susceptible S. aureus isolates. The association of the majority of isolates with SCCmec type V in one large cluster indicated the presence of a successful subclone within the clonal complex CC398 from pigs, which has diversified. In general, the combination of Cfr9I PFGE with spa and SCCmec typing demonstrated the heterogeneity of the series analyzed and can be further used for outbreak investigations and traceability studies of the MRSA ST398 emerging clone.
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the emerging methicillin resistant staphylococcus aureus st398 clone can easily be typed using the cfr9i SmaI neoschizomer
Letters in Applied Microbiology, 2010Co-Authors: M A Argudin, M R Rodicio, Beatriz GuerraAbstract:Aim: To establish a PFGE protocol using Cfr9I, neoschizomer of SmaI, for typing of Staphylococcus aureus isolates belonging to the emerging MRSA ST398 clone. Methods and Results: Staphylococcus aureus ST398 and non-ST398 isolates were analysed using the PFGE conditions recommended by the HARMONY consensus protocol. Genomic DNA of non-ST398 isolates could be digested with SmaI, XmaI (also a SmaI-neoschizomer) and Cfr9I. The DNA of SmaI-nontypeable ST398 isolates was partially resistant to XmaI, but could be digested with Cfr9I. By PCR-amplification/sequencing, the presence of a novel C5-cytosine methyltransferase gene (sauST398M) was detected in the ST398 isolates. The encoded enzyme, which shows high similarity with C5-cytosine methyltransferases that modify the CCCGGG recognition sequence, could be responsible for the different restriction results. Conclusion: SmaI-PFGE is regarded as the ‘gold standard’ for typing S. aureus. Because of different susceptibility of the GGGCCC recognition sites of the ST398 DNA against SmaI, XmaI and Cfr9I, the proposed protocol is a valuable tool for ST398 typing. Significance and Impact of the Study: The use of this protocol allows the comparison of results from SmaI-nontypeable isolates with S. aureus SmaI-PFGE databases and can be applied for outbreak investigations and traceability studies of this emerging MRSA clone.
Birgit Strommenger - One of the best experts on this subject based on the ideXlab platform.
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methicillin resistant staphylococcus aureus and staphylococcus pseudintermedius detected in the bft germvet monitoring programme 2004 2006 in germany
Journal of Antimicrobial Chemotherapy, 2007Co-Authors: Stefan Schwarz, Kristina Kadlec, Birgit StrommengerAbstract:Objectives: During recent years, methicillin-resistant Staphylococcus aureus (MRSA) strains from animals have become the focus of various studies. In the present study, coagulase-positive staphylococci obtained from pigs, dogs and cats suffering from acute infections, which had been collected in the BfT-GermVet monitoring programme in Germany and phenotypically identified as oxacillin-resistant, were characterized. Methods: The staphylococci were comparatively investigated for their resistance phenotypes and genotypes. Resistance genes were identified by PCR. MRSA strains were further characterized by SmaI macrorestriction analysis and spa typing to assess their genomic relationships. Results: Among the 248 strains tested, 7 strains (5 porcine S. aureus and 2 canine Staphylococcus pseudintermedius) carried the resistance gene mecA. Gentamicin resistance was based on the presence of the gene aacA/aphD while three different tetracycline resistance genes, tet(K), tet(L) and tet(M), alone or in combinations, were detected. The single macrolide/lincosamide-resistant strain carried an erm(A) gene. All MRSA strains proved to be non-typeable by SmaI macrorestriction analysis and exhibited the spa types t011 (four strains) or t034 (one strain). Conclusions: Based on their spa types and their non-respondence to SmaI digestion, all five porcine MRSA strains resembled MRSA strains of multilocus sequence type ST398, previously detected among pigs in neighbouring countries such as The Netherlands or Denmark. The results of this study indicate that such strains are also involved in defined disease conditions of pigs from various parts of Germany.
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assignment of staphylococcus isolates to groups by spa typing SmaI macrorestriction analysis and multilocus sequence typing
Journal of Clinical Microbiology, 2006Co-Authors: Birgit Strommenger, Christiane Kettlitz, Thomas Weniger, Dag Harmsen, Alexander W Friedrich, W WitteAbstract:The implementation of the new clustering algorithm Based Upon Repeat Pattern (BURP) into the Ridom StaphType software tool enables clustering based on spa typing data for Staphylococcus aureus. We compared clustering results obtained by spa typing/BURP to those obtained by currently well-established methods, i.e., SmaI macrorestriction analysis and multilocus sequence typing/eBURST. A total of 99 clinical S. aureus strains, including MRSA and representing major clonal lineages associated with important kinds of infections which have been prevalent in Germany and Central Europe during the last 10 years, were used for comparison. SmaI macrorestriction analysis revealed the highest discriminatory power, and clustering results for all three methods resulted in concordance values ranging from 96.8% between the two sequence-based methods to 93.4% between spa typing/BURP and SmaI macrorestriction/cluster analysis. The results of this study indicate that spa typing, together with BURP clustering, is a useful tool in S. aureus epidemiology, especially because of ease of use and the advantages of unambiguous sequence analysis as well as reproducibility and exchange of typing data.
