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2.
J Hosp Infect ; 132: 8-19, 2023 Feb.
Article in English | MEDLINE | ID: mdl-36481685

ABSTRACT

BACKGROUND: A novel Panton-Valentine leukocidin (PVL)-positive meticillin-resistant Staphylococcus aureus (MRSA) clonal complex (CC)5-MRSA-IVc ('Sri Lankan' clone) was recently described from Sri Lanka. Similar isolates caused a recent Irish hospital outbreak. AIM: To investigate the international dissemination and diversity of PVL-positive CC5-MRSA-IVc isolates from hospital and community settings using whole-genome sequencing (WGS). METHODS: Core-genome single nucleotide polymorphism (cgSNP) analysis, core-genome multi-locus sequence typing (cgMLST) and microarray-based detection of antimicrobial-resistance and virulence genes were used to investigate PVL-positive CC5-MRSA-IVc (N = 214 including 46 'Sri Lankan' clone) from hospital and community settings in 12 countries over 17 years. Comparators included 29 PVL-positive and 23 PVL-negative CC5/ST5-MRSA-I/II/IVa/IVc/IVg/V. RESULTS: Maximum-likelihood cgSNP analysis grouped 209/214 (97.7%) CC5-MRSA-IVc into Clade I; average of 110 cgSNPs between isolates. Clade III contained the five remaining CC5-MRSA-IVc; average of 92 cgSNPs between isolates. Clade II contained seven PVL-positive CC5-MRSA-IVa comparators, whereas the remaining 45 comparators formed an outlier group. Minimum-spanning cgMLST analysis revealed a comparably low average of 57 allelic differences between all CC5/ST5-MRSA-IVc. All 214 CC5/ST5-MRSA-IVc were identified as 'Sri Lankan' clone, predominantly spa type t002 (186/214) with low population diversity and harboured a similar range of virulence genes and variable antimicrobial-resistance genes. All 214 Sri Lankan clone isolates and Clade II comparators harboured a 9616-bp chromosomal PVL-encoding phage remnant, suggesting both arose from a PVL-positive meticillin-susceptible ancestor. Over half of Sri Lankan clone isolates were from infections (142/214), and where detailed metadata were available (168/214), most were community associated (85/168). CONCLUSIONS: Stable chromosomal retention of pvl may facilitate Sri-Lankan clone dissemination.


Subject(s)
Methicillin-Resistant Staphylococcus aureus , Staphylococcal Infections , Humans , Methicillin , Multilocus Sequence Typing , Staphylococcal Infections/epidemiology , Exotoxins/genetics , Leukocidins/genetics , Hospitals , Microbial Sensitivity Tests
3.
Zoonoses Public Health ; 69(6): 673-681, 2022 09.
Article in English | MEDLINE | ID: mdl-35546073

ABSTRACT

While previous research on zoonotic transmission of community-acquired Clostridioides difficile infection (CA-CDI) focused on food-producing animals, the present study aimed to investigate whether dogs are carriers of resistant and/or virulent C. difficile strains. Rectal swabs were collected from 323 dogs and 38 C. difficile isolates (11.8%) were obtained. Isolates were characterized by antimicrobial susceptibility testing, whole-genome sequencing (WGS) and a DNA hybridization assay. Multilocus sequence typing (MLST), core genome MLST (cgMLST) and screening for virulence and antimicrobial resistance genes were performed based on WGS. Minimum inhibitory concentrations for erythromycin, clindamycin, tetracycline, vancomycin and metronidazole were determined by E-test. Out of 38 C. difficile isolates, 28 (73.7%) carried genes for toxins. The majority of isolates belonged to MLST sequence types (STs) of clade I and one to clade V. Several isolates belonged to STs previously associated with human CA-CDI. However, cgMLST showed low genetic relatedness between the isolates of this study and C. difficile strains isolated from humans in Austria for which genome sequences were publicly available. Four isolates (10.5%) displayed resistance to three of the tested antimicrobial agents. Isolates exhibited resistance to erythromycin, clindamycin, tetracycline and metronidazole. These phenotypic resistances were supported by the presence of the resistance genes erm(B), cfr(C) and tet(M). All isolates were susceptible to vancomycin. Our results indicate that dogs may carry virulent and antimicrobial-resistant C. difficile strains.


Subject(s)
Anti-Infective Agents , Clostridioides difficile , Clostridium Infections , Dog Diseases , Animals , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Anti-Infective Agents/pharmacology , Clindamycin/pharmacology , Clostridioides , Clostridioides difficile/genetics , Clostridium Infections/epidemiology , Clostridium Infections/veterinary , Dog Diseases/drug therapy , Dog Diseases/epidemiology , Dogs , Drug Resistance, Bacterial/genetics , Erythromycin , Genotype , Humans , Metronidazole/pharmacology , Microbial Sensitivity Tests/veterinary , Multilocus Sequence Typing/veterinary , Tetracyclines , Vancomycin/pharmacology
5.
Vet Microbiol ; 254: 109005, 2021 Mar.
Article in English | MEDLINE | ID: mdl-33582485

ABSTRACT

Information about livestock carrying methicillin-resistant coagulase-negative staphylococci and mammaliicocci (MRCoNS/MRM) is scarce. The study was designed to gain knowledge of the prevalence, the phenotypic and genotypic antimicrobial resistance and the genetic diversity of MRCoNS/MRM originating from ruminants and New World camelids. In addition, a multi-locus sequence typing scheme for the characterization of Mammaliicoccus (formerly Staphylococcus) sciuri was developed. The study was conducted from April 2014 to January 2017 at the University Clinic for Ruminants and the Institute of Microbiology at the University of Veterinary Medicine Vienna. Seven hundred twenty-three nasal swabs originating from ruminants and New World camelids with and without clinical signs were examined. After isolation, MRCoNS/MRM were identified by MALDI-TOF, rpoB sequencing and typed by DNA microarray-based analysis and PCR. Antimicrobial susceptibility testing was conducted by agar disk diffusion. From all 723 nasal swabs, 189 MRCoNS/MRM were obtained. Members of the Mammaliicoccus (M.) sciuri group were predominant (M. sciuri (n = 130), followed by M. lentus (n = 43), M. fleurettii (n = 11)). In total, 158 out of 189 isolates showed phenotypically a multi-resistance profile. A seven-loci multi-locus sequence typing scheme for M. sciuri was developed. The scheme includes the analysis of internal segments of the house-keeping genes ack, aroE, ftsZ, glpK, gmk, pta1 and tpiA. In total, 28 different sequence types (STs) were identified among 92 selected M. sciuri isolates. ST1 was the most prevalent ST (n = 35), followed by ST 2 (n = 15), ST3 and ST5 (each n = 5), ST4 (n = 3), ST6, ST7, ST8, ST9, ST10 and ST11 (each n = 2).


Subject(s)
Camelids, New World/microbiology , Genetic Variation , Methicillin Resistance , Ruminants/microbiology , Staphylococcal Infections/veterinary , Staphylococcus/classification , Staphylococcus/genetics , Animals , Anti-Bacterial Agents/pharmacology , Bacterial Typing Techniques/methods , DNA, Bacterial/genetics , Drug Resistance, Multiple, Bacterial/genetics , Genes, Bacterial/genetics , Microbial Sensitivity Tests , Multilocus Sequence Typing/methods , Staphylococcal Infections/epidemiology , Staphylococcus/drug effects
6.
Sci Adv ; 7(5)2021 01.
Article in English | MEDLINE | ID: mdl-33571128

ABSTRACT

Many species synchronize reproductive behavior with a particular phase of the lunar cycle to increase reproductive success. In humans, a lunar influence on reproductive behavior remains controversial, although the human menstrual cycle has a period close to that of the lunar cycle. Here, we analyzed long-term menstrual recordings of individual women with distinct methods for biological rhythm analysis. We show that women's menstrual cycles with a period longer than 27 days were intermittently synchronous with the Moon's luminance and/or gravimetric cycles. With age and upon exposure to artificial nocturnal light, menstrual cycles shortened and lost this synchrony. We hypothesize that in ancient times, human reproductive behavior was synchronous with the Moon but that our modern lifestyles have changed reproductive physiology and behavior.

7.
J Hosp Infect ; 108: 72-80, 2021 Feb.
Article in English | MEDLINE | ID: mdl-33259881

ABSTRACT

BACKGROUND: Panton-Valentine leucocidin (PVL)-positive community-associated meticillin-resistant Staphylococcus aureus (CA-MRSA) is increasingly associated with infection outbreaks. AIM: To investigate multiple suspected PVL-positive CA-MRSA outbreaks using whole-genome sequencing (WGS). METHODS: Forty-six suspected outbreak-associated isolates from 36 individuals at three separate Irish hospitals (H1-H3) and from separate incidents involving separate families associated with H2 were investigated by whole-genome multi-locus sequence typing (wgMLST). FINDINGS: Two clusters (CH1 and CH2) consisting of 8/10 and 6/6 PVL-positive t008 ST8-MRSA-IVa isolates from H1 and H2, respectively, were identified. Within each cluster, neighbouring isolates were separated by ≤5 allelic differences; however, ≥73 allelic differences were identified between the clusters, indicating two independent outbreaks. Isolates from the H3 maternity unit formed two clusters (CH3-SCI and CH3-SCII) composed of four PVL-negative t4667 ST5-MRSA-V and 14 PVL-positive t002 ST5-MRSA-IVc isolates, respectively. Within clusters, neighbouring isolates were separated by ≤24 allelic differences, whereas both clusters were separated by 1822 allelic differences, indicating two distinct H3 outbreaks. Eight PVL-positive t127 ST1-MRSA-V+fus and three PVL-negative t267 ST97-MRSA-V+fus isolates from two distinct H2-associated families FC1 (N = 4) and FC2 (N = 7) formed three separate clusters (FC1 (t127), FC2 (t127) and FC2 (t267)). Neighbouring isolates within clusters were closely related and exhibited ≤7 allelic differences. Intrafamilial transmission was apparent, but the detection of ≥48 allelic differences between clusters indicated no interfamilial transmission. CONCLUSION: The frequent importation of PVL-positive CA-MRSA into healthcare settings, transmission and association with outbreaks is a serious ongoing concern. WGS is a highly discriminatory, informative method for deciphering such outbreaks conclusively.


Subject(s)
Community-Acquired Infections , Disease Outbreaks , Methicillin-Resistant Staphylococcus aureus , Staphylococcal Infections , Bacterial Typing Techniques , Community-Acquired Infections/epidemiology , Community-Acquired Infections/microbiology , Exotoxins , Female , Genome, Bacterial , Hospitals , Humans , Ireland/epidemiology , Leukocidins/genetics , Methicillin-Resistant Staphylococcus aureus/classification , Multilocus Sequence Typing , Pregnancy , Staphylococcal Infections/epidemiology , Whole Genome Sequencing
8.
Vet Microbiol ; 215: 77-82, 2018 Feb.
Article in English | MEDLINE | ID: mdl-29426410

ABSTRACT

The aim of this study was to determine the prevalence, the antimicrobial resistance patterns and the genetic diversity of methicillin-resistant Staphylococcus aureus (MRSA) from Austrian ruminants and New World camelids that were treated at the University of Veterinary Medicine, Vienna. Between April 2014 and January 2017, 723 nasal swabs originating from ruminants and New World camelids were examined. MRSA isolates were characterized by mecA/mecA1/mecC PCRs and by DNA microarray analysis. They were genotyped by spa typing, dru typing, MLST and MLVA. Glycopolymer fingerprinting by FTIR spectroscopy was also performed. Antimicrobial susceptibility testing was conducted by agar disk diffusion. Twelve MRSA isolates were mecA-positive, whereas three were mecC-positive. The MRSA isolates carried five different SCCmec elements, and belonged to three sequence types (ST45, ST130, ST398). The MRSA isolates displayed seven different resistance phenotypes. The present study describes for the first time mecC-carrying MRSA isolates originating from domesticated animals in Austria. More systematic studies are needed to unravel the role of ruminants and New World camelids as reservoirs for MRSA as a potential risk for zooanthropogenic transmission.


Subject(s)
Biodiversity , Camelids, New World/microbiology , Methicillin-Resistant Staphylococcus aureus/genetics , Ruminants/microbiology , Staphylococcal Infections/veterinary , Animals , Anti-Infective Agents/pharmacology , Austria , DNA, Bacterial/genetics , Genotype , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Microbial Sensitivity Tests , Staphylococcal Infections/microbiology
9.
Eur J Clin Microbiol Infect Dis ; 36(8): 1433-1441, 2017 Aug.
Article in English | MEDLINE | ID: mdl-28421309

ABSTRACT

Shiga toxin (Stx)-producing Escherichia coli (STECs) cause non-bloody diarrhea, hemorrhagic colitis, and hemolytic uremic syndrome, and are the primary cause of acute renal failure in children worldwide. This study investigated the correlation of genetic makeup of STEC strains as revealed by DNA microarray to clinical symptoms and the duration of STEC shedding. All STEC isolated (n = 96) from patients <10 years of age in Jönköping County, Sweden from 2003 to 2015 were included. Isolates were characterized by DNA microarray, including almost 280 genes. Clinical data were collected through a questionnaire and by reviewing medical records. Of the 96 virulence genes (including stx) in the microarray, 62 genes were present in at least one isolate. Statistically significant differences in prevalence were observed for 21 genes when comparing patients with bloody diarrhea (BD) and with non-bloody stool (18 of 21 associated with BD). Most genes encode toxins (e.g., stx2 alleles, astA, toxB), adhesion factors (i.e. espB_O157, tir, eae), or secretion factors (e.g., espA, espF, espJ, etpD, nleA, nleB, nleC, tccP). Seven genes were associated with prolonged stx shedding; the presence of three genes (lpfA, senB, and stx1) and the absence of four genes (espB_O157, espF, astA, and intI1). We found STEC genes that might predict severe disease outcome already at diagnosis. This can be used to develop diagnostic tools for risk assessment of disease outcome. Furthermore, genes associated with the duration of stx shedding were detected, enabling a possible better prediction of length of STEC carriage after infection.


Subject(s)
Bacterial Shedding , Escherichia coli Infections/microbiology , Escherichia coli Infections/pathology , Microarray Analysis , Shiga-Toxigenic Escherichia coli/classification , Shiga-Toxigenic Escherichia coli/genetics , Virulence Factors/genetics , Child , Child, Preschool , Genetic Variation , Humans , Infant , Shiga-Toxigenic Escherichia coli/isolation & purification , Sweden
10.
New Microbes New Infect ; 14: 13-8, 2016 Nov.
Article in English | MEDLINE | ID: mdl-27621823

ABSTRACT

Changes in the molecular epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) continue to be reported. This study was carried out to characterize MRSA isolates in Saudi Arabia. MRSA isolates causing nosocomial infections (n = 117) obtained from 2009-2015 at a tertiary-care facility in Riyadh, Saudi Arabia, were studied. Molecular characterization of isolates was carried out using the StaphyType DNA microarray (Alere Technologies, Jena, Germany). Fourteen clonal complexes (CC) were identified, with the most common being CC80 (n = 35), CC6 (n = 15), CC5 (n = 13) and CC22 (n = 12). With the exception of nine ST239 MRSA-III isolates, all others were of community-associated MRSA lineages. The following strains are identified for the first time in Saudi Arabia: ST8-MRSA-IV [PVL(+)/ACME(+)], USA300 (n = 1); ST72-MRSA-IV USA700 (n = 1); CC5-MRSA-IV, [PVL(+)/edinA(+)], WA MRSA-121 (n = 1); CC5-MRSA-V+SCCfus, WA MRSA-14/109 (n = 2), CC97-MRSA-IV, WA MRSA-54/63; CC2250/2277-MRSA-IV and WA MRSA-114. CC15-MRSA (n = 3) was identified for the first time in clinical infection in Saudi Arabia. None of the isolates harboured vancomycin resistance genes, while genes for resistance to mupirocin and quaternary ammonium compounds were found in one and nine isolates respectively. Fifty-seven isolates (48.7%) were positive for Panton-Valentine leukocidin genes. While the staphylokinase (sak) and staphylococcal complement inhibitor (scn) genes were present in over 95% of the isolates, only 37.6% had the chemotaxis-inhibiting protein (chp) gene. Increasing occurrence of community-acquired MRSA lineages plus emergence of pandemic and rare MRSA strains is occurring in our setting. Strict infection control practices are important to limit the dissemination of these MRSA strains.

11.
Eur J Clin Microbiol Infect Dis ; 35(12): 1963-1973, 2016 Dec.
Article in English | MEDLINE | ID: mdl-27553495

ABSTRACT

While there is an abundance of data on the epidemiology and molecular typing of Staphylococcus aureus, especially those carrying Panton-Valentine leucocidin (PVL) genes or mecA from Western Europe, Northern America and Australia, comparably few studies target African strains. In this study, we characterised genes associated with virulence and resistance, as well the phylogenetic background of S. aureus from healthy carriers and outpatients in Gabon. In total, 103 isolates from 96 study participants were characterised. Seventy-nine isolates originated from throat swabs and 24 isolates from skin lesions. Three isolates carried mecA, although only one, belonging to CC8-MRSA-IV [PVL+] 'USA300', was found to be phenotypically oxacillin-resistant; two CC88-MRSA-IV isolates appeared to be oxacillin-susceptible. PVL genes were common, with a total of 44 isolates (43 %) found to be PVL-positive. CC15-MSSA [PVL+] (n = 29) and CC152-MSSA [PVL+] (n = 9) were the predominant clones among the PVL-positive isolates. Among PVL-negative isolates, CC5-MSSA (n = 12), CC101-MSSA (n = 10) and CC15 (n = 9) were the most frequent. A hitherto undescribed multilocus sequence type of S. schweitzeri was detected twice in unrelated patients. The data emphasise a need for further studies on the role of PVL in African populations and the clinical significance of S. schweitzeri.


Subject(s)
Staphylococcal Infections/epidemiology , Staphylococcus aureus/classification , Staphylococcus aureus/genetics , Adolescent , Adult , Bacterial Proteins/genetics , Bacterial Toxins/genetics , Carrier State/epidemiology , Carrier State/microbiology , Child , Child, Preschool , Cross-Sectional Studies , Exotoxins/genetics , Female , Gabon/epidemiology , Genotype , Humans , Leukocidins/genetics , Male , Molecular Epidemiology , Multilocus Sequence Typing , Outpatients , Penicillin-Binding Proteins/genetics , Pharynx/microbiology , Skin/microbiology , Staphylococcal Infections/microbiology , Staphylococcus aureus/isolation & purification
12.
Eur J Clin Microbiol Infect Dis ; 35(9): 1541-7, 2016 Sep.
Article in English | MEDLINE | ID: mdl-27262852

ABSTRACT

Comparatively few studies have been published describing Staphylococcus aureus/MRSA epidemiology in Central Asia including Pakistan. Here, we report the genotyping of Staphylococcus aureus strains (that include both methicillin-susceptible and methicillin-resistant Staphylococcus aureus) from community- and hospital-acquired skin and soft-tissue infections in a tertiary care hospital in the Malakand district of the Khyber Pakhtunkhwa Province of Pakistan. Forty-five isolates of Staphylococcus aureus were characterized by microarray hybridization. Twenty isolates (44 %) were MRSA, whereas 22 (49 %) were PVL-positive. Fourteen isolates (31 %) harboured both mecA and PVL genes. The dominant clones were CC121-MSSA (n = 15, 33 %) and the PVL-positive "Bengal Bay Clone" (ST772-MRSA-V; n = 13, 29 %). The PVL-positive CC8-MRSA-IV strain "USA300" was found once. The pandemic ST239-MRSA-III strain was absent, although it has previously been observed in Pakistan. These observations require a re-assessment of schemes for initial antibiotic therapy to cover MRSA and they emphasise the need for a rapid and non-molecular test for PVL.


Subject(s)
Soft Tissue Infections/microbiology , Staphylococcal Skin Infections/microbiology , Staphylococcus aureus/classification , Staphylococcus aureus/genetics , Bacterial Proteins/genetics , Bacterial Toxins/genetics , Community-Acquired Infections/epidemiology , Community-Acquired Infections/microbiology , Cross Infection/epidemiology , Cross Infection/microbiology , Exotoxins/genetics , Genotype , Genotyping Techniques , Humans , Leukocidins/genetics , Microarray Analysis , Molecular Epidemiology , Nucleic Acid Hybridization , Pakistan/epidemiology , Penicillin-Binding Proteins/genetics , Soft Tissue Infections/epidemiology , Staphylococcal Skin Infections/epidemiology , Staphylococcus aureus/isolation & purification , Tertiary Care Centers
13.
Eur J Clin Microbiol Infect Dis ; 35(8): 1285-95, 2016 Aug.
Article in English | MEDLINE | ID: mdl-27177754

ABSTRACT

Providing evidence for microbial genetic determinants' impact on outcome in Staphylococcus aureus bloodstream infections (SABSI) is challenging due to the complex and dynamic microbe-host interaction. Our recent population-based prospective study reported an association between the S. aureus clonal complex (CC) 30 genotype and mortality in SABSI patients. This follow-up investigation aimed to examine the genetic profiles of the SABSI isolates and test the hypothesis that specific genetic characteristics in S. aureus are associated with mortality. SABSI isolates (n = 305) and S. aureus CC30 isolates from asymptomatic nasal carriers (n = 38) were characterised by DNA microarray analysis and spa typing. Fisher's exact test, least absolute shrinkage and selection operator (LASSO) and elastic net regressions were performed to discern within four groups defined by patient outcome and characteristics. No specific S. aureus genetic determinants were found to be associated with mortality in SABSI patients. By applying LASSO and elastic net regressions, we found evidence suggesting that agrIII and cna were positively and setC (=selX) and seh were negatively associated with S. aureus CC30 versus non-CC30 isolates. The genes chp and sak, encoding immune evasion molecules, were found in higher frequencies in CC30 SABSI isolates compared to CC30 carrier isolates, indicating a higher virulence potential. In conclusion, no specific S. aureus genes were found to be associated with mortality by DNA microarray analysis and state-of-the-art statistical analyses. The next natural step is to test the hypothesis in larger samples with higher resolution methods, like whole genome sequencing.


Subject(s)
Bacteremia , Host-Pathogen Interactions/genetics , Staphylococcal Infections , Staphylococcus aureus , Aged , Aged, 80 and over , Bacteremia/microbiology , Bacteremia/mortality , DNA, Bacterial/analysis , DNA, Bacterial/genetics , Female , Genes, Bacterial/genetics , Humans , Male , Microarray Analysis , Middle Aged , Prospective Studies , Staphylococcal Infections/microbiology , Staphylococcal Infections/mortality , Staphylococcus aureus/genetics , Staphylococcus aureus/pathogenicity
14.
Eur J Clin Microbiol Infect Dis ; 35(8): 1355-61, 2016 Aug.
Article in English | MEDLINE | ID: mdl-27193891

ABSTRACT

Knowledge on Staphylococcus aureus colonization rates and epidemiology in hand eczema is limited. The aim of this study was to clarify some of these issues. Samples were collected by the "glove juice" method from the hands of 59 patients with chronic hand eczema and 24 healthy individuals. Swab samples were taken from anterior nares and throat from 43 of the 59 patients and all healthy individuals. S. aureus were spa typed and analysed by DNA-microarray-based genotyping. The extent of the eczema was evaluated by the hand eczema extent score (HEES). The colonization rate was higher on the hands of hand eczema patients (69 %) compared to healthy individuals (21 %, p < 0.001). This was also seen for bacterial density (p = 0.002). Patients with severe hand eczema (HEES ≥ 13) had a significantly higher S. aureus density on their hands compared to those with milder eczema (HEES = 1 to 12, p = 0.004). There was no difference between patients and healthy individuals regarding colonization rates in anterior nares or throat. spa typing and DNA-microarray-based genotyping indicated certain types more prone to colonize eczematous skin. Simultaneous colonization, in one individual, with S. aureus of different types, was identified in 60-85 % of the study subjects. The colonization rate and density indicate a need for effective treatment of eczema and may have an impact on infection control in healthcare.


Subject(s)
Eczema , Staphylococcal Infections , Staphylococcal Skin Infections , Staphylococcus aureus/isolation & purification , Anti-Bacterial Agents/pharmacology , Case-Control Studies , Drug Resistance, Bacterial , Eczema/complications , Eczema/microbiology , Female , Humans , Male , Molecular Typing , Staphylococcal Infections/complications , Staphylococcal Infections/microbiology , Staphylococcal Skin Infections/complications , Staphylococcal Skin Infections/microbiology , Staphylococcus aureus/drug effects , Staphylococcus aureus/genetics
15.
Clin Microbiol Infect ; 21(5): 444-50, 2015 May.
Article in English | MEDLINE | ID: mdl-25708549

ABSTRACT

Typing of healthcare-associated methicillin-resistant Staphylococcus aureus (MRSA) from Australia in the 1970s revealed a novel clone, ST2249-MRSA-III (CC45), present from 1973 to 1979. This clone was present before the Australian epidemic caused by the recombinant clone, ST239-MRSA-III. This study aimed to characterize the genome of ST2249-MRSA-III to establish its relationship to other MRSA clones. DNA microarray analysis was conducted and a draft genome sequence of ST2249 was obtained. The recombinant structure of the ST2249 genome was revealed by comparisons to publicly available ST239 and ST45 genomes. Microarray analysis of genomic DNA of 13 ST2249 isolates showed gross similarities with the ST239 chromosome in a segment around the origin of replication and with ST45 for the remainder of the chromosome. Recombination breakpoints were precisely determined by the changing pattern of nucleotide polymorphisms in the genome sequence of ST2249 isolate SK1585 compared with ST239 and ST45. One breakpoint was identified to the right of oriC, between sites 1014 and 1065 of the gene D484_00045. Another was identified to the left of oriC, between sites 1185 and 1248 of D484_01632. These results indicate that ST2249 inherited approximately 35.3% of its chromosome from an ST239-like parent and 64.7% from an ST45-like parent. ST2249-MRSA-III resulted from a major recombination between parents that resemble ST239 and ST45. Although only limited Australian archival material is available, the oldest extant isolate of ST2249 predates the oldest Australian isolate of ST239 by 3 years. It is therefore plausible that these two recombinant clones were introduced into Australia separately.


Subject(s)
Cross Infection/epidemiology , Cross Infection/microbiology , Genotype , Methicillin-Resistant Staphylococcus aureus/classification , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Recombination, Genetic , Staphylococcal Infections/epidemiology , Australia/epidemiology , Chromosomes, Bacterial , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Evolution, Molecular , Genome, Bacterial , Humans , Methicillin-Resistant Staphylococcus aureus/genetics , Microarray Analysis , Sequence Analysis, DNA , Staphylococcal Infections/microbiology
16.
Vet Microbiol ; 176(1-2): 143-54, 2015 Mar 23.
Article in English | MEDLINE | ID: mdl-25631254

ABSTRACT

Staphylococcus aureus is one of the most relevant pathogens causing clinical and subclinical, chronic mastitis in dairy animals. Routinely, mastitis pathogens are isolated and classified to genus or species level, and regarded as single entities. However, S. aureus includes a broad range of genotypes with distinct pathogenic and epidemiologic characteristics. The objective of the present study was to assess the host-specificity of S. aureus causing mastitis in dairy animals, based on phylogenetic and genotypic characterization as well as the presence of virulence and antimicrobial resistance genes in the pathogen genome. S. aureus isolates from mastitis in cows, sheep and goats in Israel, and from cows in Germany, the USA and Italy, were compared by the following methods: a. Bayesian phylogenetic comparison of sequences of genes nuc, coa, lukF and clfA, b. genotyping by spa and agr typing, and assignment to MLST Clonal Complexes (MLST CC), and c. the presence of a broad array of virulence and antimicrobial resistance genes. Overall, phylogenetic, virulence and genotyping approaches agreed with each other. Cow isolates could be differentiated from sheep and goat isolates with all three methods, with different resolution. In two phylogenetic clusters, segregation was found also between cow isolates from Israel and abroad. Sheep and goats' isolates showed less variability than isolates from cows in all methods used. In conclusion, different S. aureus lineages are associated to cows in contrast to goats and sheep, suggesting co-evolution between pathogen and host species. Modern diagnostics approaches should aim to explore molecular data for a better understanding and cost-effective management of mastitis.


Subject(s)
Cattle Diseases/microbiology , Goat Diseases/microbiology , Mastitis/veterinary , Staphylococcal Infections/veterinary , Staphylococcus aureus/metabolism , Virulence Factors/metabolism , Animals , Cattle , Female , Genotype , Germany , Goats/genetics , Host Specificity , Israel , Italy , Mastitis/microbiology , Multilocus Sequence Typing , Phylogeny , Sheep/genetics , Sheep Diseases/microbiology , Staphylococcus aureus/classification , Staphylococcus aureus/genetics , Virulence/genetics , Virulence Factors/genetics
17.
J Antimicrob Chemother ; 70(3): 649-52, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25411186

ABSTRACT

OBJECTIVES: To describe a family of conjugative plasmids isolated from colonizing community Staphylococcus aureus and determine their ability to mobilize unrelated antimicrobial resistance/virulence plasmids, not encoding mobilization functions. METHODS: Plasmid pWBG749 was labelled with Tn551 (pWBG749e) to enable laboratory manipulation. Plasmid pWBG749e was conjugated into S. aureus of seven different lineages that harboured unrelated plasmids and mobilization experiments were performed. Plasmids were screened by EcoRI restriction and hybridization with probes prepared from unique pWBG749 conjugation genes. RESULTS: Conjugative plasmids pWBG745, pWBG748 and pWBG749 belong to the same conjugative-plasmid family as the vancomycin resistance plasmid pBRZ01. Plasmid pWBG749e mobilized five unrelated plasmids. Mobilized plasmid pWBG744 is a pIB485-family plasmid that was also found in international S. aureus. CONCLUSIONS: Plasmid pWBG749e can mobilize unrelated S. aureus plasmids whose means of dissemination have not previously been understood.


Subject(s)
Conjugation, Genetic , Gene Transfer, Horizontal , Plasmids , Staphylococcus aureus/genetics , Community-Acquired Infections/microbiology , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Gene Order , Genes, Bacterial , Humans , Molecular Sequence Data , Sequence Analysis, DNA , Staphylococcal Infections/microbiology , Staphylococcus aureus/isolation & purification
18.
J Dairy Sci ; 97(5): 2782-8, 2014 May.
Article in English | MEDLINE | ID: mdl-24630668

ABSTRACT

The discovery of a new mecA homolog, mecC, necessitates a modification of diagnostic procedures for the identification of methicillin-resistant Staphylococcus aureus (MRSA), as most assays used for the genotypic and phenotypic mecA detection cannot currently recognize mecC. Although the prevalence, distribution, and importance of mecC are not yet completely understood, an exchange of mecC-MRSA between humans and animals seems possible. All previously reported observations of mecC-positive strains have been sporadic. To the best of our knowledge, this is the first report about multiple cases of mecC-positive Staph. aureus in 1 dairy herd. Clonal complex 130 Staph. aureus harboring mecC were found in milk samples from 16 of 56 lactating cows kept in a herd in Bavaria, Germany. Almost all quarter milk samples positive for mecC-MRSA had the lowest possible California Mastitis Test score; composite somatic cell counts obtained from monthly milk recordings showed a mean of 51,600 cells/mL in mecC-MRSA affected cows. Additionally, mecC-positive clonal complex 130 Staph. aureus were detected in swab samples from the mammary skin and a teat lesion of 1 cow from this herd. This report suggests that mecC-carrying strains are able to spread among livestock, and that they have the ability to cause multiple cases in single herds. Therefore, future studies targeting MRSA in dairy cows need to consider mecC.


Subject(s)
Cattle Diseases/microbiology , Methicillin Resistance/genetics , Methicillin-Resistant Staphylococcus aureus/genetics , Milk/microbiology , Staphylococcal Infections/veterinary , Animals , Bacterial Proteins/genetics , Bacterial Typing Techniques/veterinary , Cattle , Cattle Diseases/epidemiology , Dairying , Female , Genotype , Germany/epidemiology , Lactation , Methicillin-Resistant Staphylococcus aureus/classification , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Oligonucleotide Array Sequence Analysis/veterinary , Skin/microbiology , Staphylococcal Infections/epidemiology , Staphylococcal Infections/microbiology
19.
Eur J Clin Microbiol Infect Dis ; 33(4): 651-8, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24162256

ABSTRACT

The purpose of this investigation was to compare the genotypic profiles of Staphylococcus aureus isolated from atopic dermatitis (AD) patients and from control subjects, and to study the relationship between clinical severity, immune response, and genomic pattern of S. aureus isolated from AD patients. We selected 32 patients with AD and S. aureus skin colonization and 31 atopic controls with no history of AD who where asymptomatic carriers of S. aureus. Microarray-based genotyping was performed on S. aureus isolates. In AD patients, clinical severity was assessed using the Scoring Atopic Dermatitis index and total IgE levels and staphylococcal superantigen-specific IgE levels (SEA, SEB, SEC, TSST1) were determined. The genes lukE, lukD, splA, splB, ssl8, and sasG were more frequent in isolates from AD patients. CC30 was more common in isolates from atopic controls than in AD patients. There was a correlation between total IgE and clinical severity, but an association between clinical severity, immune response, and the presence of S. aureus superantigen genes, including enterotoxin genes, could not be demonstrated. Finally, a correlation was found between AD severity and other S. aureus genes, such as sasG and scn. S. aureus factors besides superantigens could be related to the worsening and onset of AD.


Subject(s)
Dermatitis, Atopic/microbiology , Staphylococcal Infections/microbiology , Staphylococcus aureus/genetics , Superantigens/genetics , Adolescent , Adult , Asymptomatic Diseases , Case-Control Studies , Child , Child, Preschool , Dermatitis, Atopic/immunology , Female , Genes, Bacterial , Humans , Male , Middle Aged , Staphylococcal Infections/immunology , Staphylococcus aureus/immunology , Staphylococcus aureus/isolation & purification , Staphylococcus aureus/pathogenicity , Superantigens/immunology , Virulence/genetics , Young Adult
20.
Appl Environ Microbiol ; 79(2): 655-62, 2013 Jan.
Article in English | MEDLINE | ID: mdl-23160118

ABSTRACT

Staphylococcus aureus causes a wide range of infectious diseases in humans and various animal species. Although presumptive host-specific factors have been reported, certain genetic lineages seem to lack specific host tropism, infecting a broad range of hosts. Such Extended-Host-Spectrum Genotypes (EHSGs) have been described in canine infections, caused by common regional human methicillin-resistant S. aureus (MRSA) lineages. However, information is scarce about the occurrence of methicillin-susceptible S. aureus (MSSA) EHSGs. To gain deeper insight into EHSG MSSA and EHSG MRSA of human and canine origin, a comparative molecular study was carried out, including a convenience sample of 120 current S. aureus (70 MRSA and 50 MSSA) isolates obtained from infected dogs. spa typing revealed 48 different spa types belonging to 16 different multilocus sequence typing clonal complexes (MLST-CCs). Based on these results, we further compared a subset of canine (n = 48) and human (n = 14) strains, including isolates of clonal complexes CC5, CC22, CC8, CC398, CC15, CC45, and CC30 by macrorestriction (pulsed-field gel electrophoresis [PFGE]) and DNA-microarray analysis. None of the methods employed was able to differentiate between clusters of human and canine strains independently of their methicillin resistance. In contrast, DNA-microarray analysis revealed 79% of the 48 canine isolates as carriers of the bacteriophage-encoded human-specific immune evasion cluster (IEC). In conclusion, the high degree of similarity between human and canine S. aureus strains regardless of whether they are MRSA or MSSA envisions the existence of common genetic traits that enable these strains as EHSGs, challenging the concept of resistance-driven spillover of MRSA.


Subject(s)
Staphylococcal Infections/microbiology , Staphylococcal Infections/veterinary , Staphylococcus aureus/classification , Staphylococcus aureus/isolation & purification , Animals , Bacteriophages/genetics , Cluster Analysis , Dogs , Electrophoresis, Gel, Pulsed-Field , Genotype , Host Specificity , Humans , Methicillin Resistance , Microarray Analysis , Multilocus Sequence Typing , Staphylococcal Infections/epidemiology , Staphylococcus aureus/genetics , Staphylococcus aureus/physiology
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