Clinical and molecular features of methicillin-resistant,coagulase-negative staphylococci of pets and horses

Objectives To determine the antibiotic resistance and fingerprint profiles of methicillin-resistant coagulase-negative staphylococci (MRCoNS) from animal infections among different practices and examine the history of antibiotic treatment. Methods Isolates were identified by mass spectrometry and tested for antimicrobial resistance by broth dilution, microarrays and sequence analysis of the topoisomerases. Diversity was assessed by PFGE, icaA PCR and staphylococcal cassette chromosome mec (SCCmec), arginine catabolic mobile element (ACME) and multilocus sequence typing. Clinical records were examined retrospectively. Results MRCoNS were identified as Staphylococcus epidermidis (n = 20), Staphylococcus haemolyticus (n = 17), Staphylococcus hominis (n = 3), Staphylococcus capitis (n = 1), Staphylococcus cohnii (n = 1) and Staphylococcus warneri (n = 1). PFGE identified one clonal lineage in S. hominis isolates and several in S. haemolyticus and S. epidermidis. Fourteen sequence types were identified in S. epidermidis, with sequence type 2 (ST2) and ST5 being predominant. Ten isolates contained SCCmec IV, seven contained SCCmec V and the others were non-typeable. ACMEs were detected in 11 S. epidermidis isolates. One S. hominis and 10 S. epidermidis isolates were icaA positive. In addition to mecA-mediated β-lactam resistance, the most frequent resistance was to gentamicin/kanamycin [aac(6′)-Ie-aph(2′)-Ia, aph(3′)-III] (n = 34), macrolides/lincosamides [erm(C), erm(A), msr, lnu(A)] (n = 31), tetracycline [tet(K)] (n = 22), streptomycin [str, ant(6)-Ia] (n = 20), trimethoprim [dfr(A), dfr(G)] (n = 17), sulfamethoxazole (n = 34) and fluoroquinolones [amino acid substitutions in GyrA and GrlA] (n = 30). Clinical data suggest selection through multiple antibiotic courses and emphasize the importance of accurate diagnosis and antibiograms. Conclusions MRCoNS from animal infection sites are genetically heterogeneous multidrug-resistant strains that represent a new challenge in the prevention and therapy of infections in veterinary clinic

Whole-Genome Sequences of Antibiotic-Resistant Trueperella pyogenes Isolates from Surgical Site Infections in Dairy Cows in Switzerland.

The complete genome sequence of four Trueperella pyogenes isolated from cattle surgical site infections in Switzerland was determined using hybrid assembly of Oxford Nanopore and Illumina reads. Genes conferring resistance to tetracyclines [tet(W)], sulfonamides (sul1), chloramphenicol (cmx), streptomycin/spectinomycin (aadA1), and quaternary ammonium compounds (qacEΔ1) were identified on different chromosomal elements

Novel macrolide-lincosamide-streptogramin B resistance gene erm(56) in Trueperella pyogenes

Suppurative infections caused by Trueperella pyogenes, a commensal and opportunistic Gram-positive pathogen of animals, are occasionally treated using macrolides and lincosamides posing the risk of antimicrobial resistance selection. Acquired resistances to macrolide, lincosamide and streptogramin B (MLSB) antibiotics in T. pyogenes have been so far associated with erythromycin ribosome methylase genes, erm(B) or erm(X), located within mobile genetic elements. T. pyogenes strain 09KM1269, isolated from a dog abscess, exhibited constitutive resistance to erythromycin and clindamycin. Whole genome sequence analysis identified a novel gene, erm(56), that coded for a 23S rRNA methylase and showed the closest relatedness to Erm(X) with only 54% nucleotide and 58% amino acid identity. Functionality of the new gene was demonstrated by cloning erm(56) and its promoter sequences into pJRD215. The resulting erm(56)-containing plasmid pJEM1269 was subsequently electrotransformed into susceptible strains of E. coli AG100A and T. pyogenes 13OD0707. When erm(56) was expressed from pJEM1269 in T. pyogenes 13OD0707, the MIC increased by more than 256-fold for erythromycin and clindamycin and by 16-fold for pristinamycin IA. Increased MICs of erythromycin (64-fold) and clindamycin (8-fold) were also measured for E. coli AG100A containing pJEM1269. The erm(56) gene was integrated into the chromosome between two IS6100, situated next to a class 1 integron containing the sulfonamide resistance gene sul1. The erm(56) gene associated with IS6100 was also detected in strains from livestock in China, namely in another T. pyogenes and a Rothia nasimurium. Although a circular conformation containing one copy of IS6100 was detected by PCR, the erm(56) gene could not be transferred by either filter mating or electroporation of genomic DNA into MLSB-susceptible and plasmid-free T. pyogenes strains. The detection of erm(56) in unrelated bacteria from different animal sources and geographical origins suggests that it has been independently acquired and likely selected by the use of antibiotics

Novel macrolide-lincosamide-streptogramin B resistance gene erm(56) in Trueperella pyogenes.

Whole-genome sequence analysis of a macrolide, lincosamide, streptogramin B (MLSB)-resistant Trueperella pyogenes from a dog revealed a new 23S ribosomal RNA methylase gene erm(56). Expression of the cloned erm(56) confers resistance to MLSB in T. pyogenes and Escherichia coli. The erm(56) gene was flanked by two IS6100 integrated on the chromosome next to a sul1-containing class 1 integron. GenBank query revealed additional erm(56)-containing elements in another T. pyogenes and in Rothia nasimurium from livestock. IMPORTANCE A novel 23S ribosomal RNA methylase gene erm(56) flanked by insertion sequence IS6100 was identified in a Trueperella pyogenes isolated from the abscess of a dog and was also present in another T. pyogenes and in Rothia nasimurium from livestock. It was shown to confer resistance to macrolide, lincosamide, streptogramin B antibiotics in T. pyogenes and E. coli, indicating functionality in both Gram-positive and Gram-negative bacteria. The detection of erm(56) on different elements in unrelated bacteria from different animal sources and geographical origins suggests that it has been independently acquired and likely selected by the use of antibiotics in animals

GSFC Network Operations with Tracking and Data Relay Satellites

Since the first flights into space, the Goddard Space Flight Center has been providing various levels of support to Users of its data acquisition network. The Tracking and Data Relay Satellite System (TDRSS) Network (TN) has been developed as part of an evolving network program to continue to meet the increased communications and orbit determination needs of Users with advanced spacecraft in near-earth orbits. The extensive changes to the various network elements are nearing completion of their implementation, integration and test phases in preparation for the series of Tracking Data Relay Satellite (TDRS) launches beginning in early 1983. The final phase of integration requires extensive post-flight testing of the TDRS\u27s with operational network elements before the TN can be declared operational for full Users support. The TDRSS Network is unique from past network development concepts in that it comprises a combination of comirencal and government elements integrated into a highly automated end-to-end system!. The commercial portion, a series of satellites In geostationary orbit monitored and controlled by a central ground terminal is intended to replace NASA\u27s present geographically distributed ground terminals. The NASA portion comprises a series of elements that provide unique services and support to monitor and control the entire network under a mission support type contract. An overview of this system that is planned to fulfill the Users scientific requirements in the 80\u27s and into the 90\u27s is presented

Comparative genomics of 26 complete circular genomes of 18 different serotypes of Actinobacillus pleuropneumoniae.

Actinobacillus pleuropneumoniae is a Gram-negative, rod-shaped bacterium of the family Pasteurellaceae causing pig pleuropneumonia associated with great economic losses worldwide. Nineteen serotypes with distinctive lipopolysaccharide (LPS) and capsular (CPS) compositions have been described so far, yet complete circular genomes are publicly available only for the reference strains of serotypes 1, 4 and 5b, and for field strains of serotypes 1, 3, 7 and 8. We aimed to complete this picture by sequencing the reference strains of 17 different serotypes with the MinION sequencer (Oxford Nanopore Technologies, ONT) and on an Illumina HiSeq (Illumina) platform. We also included two field isolates of serotypes 2 and 3 that were PacBio- and MinION-sequenced, respectively. Genome assemblies were performed following two different strategies, i.e. PacBio- or ONT-only de novo assemblies polished with Illumina reads or a hybrid assembly by directly combining ONT and Illumina reads. Both methods proved successful in obtaining accurate circular genomes with comparable qualities. blast-based genome comparisons and core-genome phylogeny based on core genes, SNP typing and multi-locus sequence typing (cgMLST) of the 26 circular genomes indicated well-conserved genomes across the 18 different serotypes, differing mainly in phage insertions, and CPS, LPS and RTX-toxin clusters, which, consistently, encode serotype-specific antigens. We also identified small antibiotic resistance plasmids, and complete subtype I-F and subtype II-C CRISPR-Cas systems. Of note, highly similar clusters encoding all those serotype-specific traits were also found in other pathogenic and commensal Actinobacillus species. Taken together with the presence of transposable elements surrounding these loci, we speculate a dynamic intra- and interspecies exchange of such virulence-related factors by horizontal gene transfer. In conclusion, our comprehensive genomics analysis provides useful information for diagnostic test and vaccine development, but also for whole-genome-based epidemiological studies, as well as for the surveillance of the evolution of antibiotic resistance and virulence genes in A. pleuropneumoniae

A novel erm(44) gene variant from a human Staphylococcus saprophyticus confers resistance to macrolides, lincosamides but not streptogramins.

A novel erm (44) gene variant, erm (44)v, has been identified by whole genome sequencing in a Staphylococcus saprophyticus isolated from the skin of a healthy person. It has the particularity to confer resistance to macrolides and lincosamides, but not to streptogramins B when expressed in S. aureus The erm (44)v gene resides on a 19,400-bp genomic island which contains phage-associated proteins and is integrated into the chromosome of S. saprophyticus

The increase of methicillin-resistant Staphylococcus aureus (MRSA) and the presence of an unusual sequence type ST49 in slaughter pigs in Switzerland

<p>Abstract</p> <p>Background</p> <p>In years past, methicillin-resistant <it>S. aureus </it>(MRSA) has been frequently detected in pigs in Europe, North America and Asia. Recent, yet sporadic studies have revealed a low occurrence of MRSA in Switzerland. In 2009, a monitoring survey of the prevalence and genetic diversity of methicillin-resistant <it>S. aureus </it>(MRSA) in slaughter pigs in Switzerland was conducted using methods recommended by the EU guidelines, and using a sampling strategy evenly distributed throughout the year and representative of the Swiss slaughter pig population. Monitoring should determine if the overall prevalence of MRSA in the entire country is increasing over the years and if specific multi-resistant MRSA clones are spreading over the country.</p> <p>Results</p> <p>In 2009, the nasal cavities of eight out of 405 randomly selected pigs were positive for MRSA, representing a prevalence of 2.0% (95% CI 0.9-3.9). The following year, 23 out of 392 pigs were positive for MRSA [5.9% prevalence (95% CI 3.8-8.7)]. Three multilocus sequence types (ST), four <it>spa </it>types and two types of staphylococcal cassette chromosome <it>mec </it>(SCC<it>mec</it>) elements were detected. The most frequent genotypes were ST398 (MLST)-(<it>spa</it>)t034-V(SCC<it>mec</it>) (n = 18) and ST49-t208-V (n = 7), followed by ST398-t011-V (n = 4), ST398-t1451-V (n = 1), and ST1-t2279-IVc (n = 1). The isolates displayed resistance to ß-lactams [<it>mecA</it>, (31/31); <it>blaZ</it>, (19/31)]; tetracycline [<it>tet</it>(M), (31/31); <it>tet</it>(K), (30/31)] (n = 31); macrolides and lincosamides [<it>erm</it>(C) (4/31) or <it>erm</it>(A) (18/31)] (n = 22); tiamulin [<it>vga</it>(A)<it>v </it>(9/31) or unknown mechanism (18/31)] (n = 27); trimethoprim [<it>dfr</it>(G) (18/31); spectinomycin [<it>ant(9)-Ia </it>(19/31) or unknown mechanism (3/31)] (n = 22); streptomycin [<it>str </it>(19/31)]; sulphamethoxazole (7/31) and ciprofloxacin (n = 1) (mechanisms not determined).</p> <p>Conclusions</p> <p>This study is the first to describe the presence of MRSA ST49 in slaughter pigs, and to demonstrate a significant and nearly three-fold increase of MRSA prevalence in pigs within two years. The presence of a specific clonal lineage of MRSA from Switzerland suggests that it has been selected in Swiss pig husbandry. Effective hygiene measures should be enhanced within the entire pig production chain to suppress the spread of these pathogens into the community.</p