Microarray-Based Characterization of Staphylococcus aureus Isolates Obtained from Chicken Carcasses

2013 ◽  
Vol 76 (8) ◽  
pp. 1471-1474 ◽  
Author(s):  
REBECCA EBNER ◽  
SOPHIA JOHLER ◽  
HENNA-MARIA SIHTO ◽  
ROGER STEPHAN ◽  
CLAUDIO ZWEIFEL

A total of 34 Staphylococcus aureus isolates from flock-wise pooled chicken neck skin samples collected at two abattoirs during slaughter were characterized with DNA microarray analysis and spa typing. The 20 isolates from abattoir A all belonged to clonal complex (CC) 12 and spa type t160. Of the 14 isolates from abattoir B, 7 belonged to CC5–t3478, 5 to CC12–t160, 1 to CC45–t040, and 1 to CC101–t056. Of the various resistance-associated genes tested, only blaZ/R/I (6 isolates of CC12 and CC101 from abattoir B), sdrM (n = 34), fosB (n = 33), and qacC (n = 22) were detected. None of the isolates harbored genes conferring methicillin resistance. In terms of genes encoding enterotoxins, seb (all isolates of CC12), egc (seg, sei, selm, seln, selo, selu; all isolates of CC5 and CC45), and sea (14 isolates of CC12 and 1 isolate of CC5) were found. In addition, all isolates harbored genes for intracellular adhesion proteins (icaA/C/D) and were positive for cap5 or cap8 (capsule type 5 or 8). Comparison of DNA microarray profiles identified four categories comprising (i) all isolates of CC12, (ii) all isolates of CC5, (iii) the CC45 isolate, and (iv) the CC101 isolate. The high similarity of the isolates from abattoir A could indicate contamination of chicken carcasses with S. aureus persisting on the slaughter equipment, but further investigations are required to elucidate potential contamination routes.

2016 ◽  
Vol 145 (2) ◽  
pp. 386-396 ◽  
Author(s):  
C. HORNER ◽  
L. UTSI ◽  
L. COOLE ◽  
M. DENTON

SUMMARYWe investigated the epidemiology and characterization of isolates of Staphylococcus aureus within the Yorkshire and Humber (YH) region in the UK. In July 2015, each laboratory within YH (n = 14) was assigned two consecutive days during which all clinical isolates of S. aureus were collected. Isolates were tested for antibiotic susceptibilities and the presence of genes encoding methicillin resistance (mecA and mecC), Panton–Valentine leukocidin (PVL) (lukS-PV), and efflux-mediated chlorhexidine resistance (qacA); isolates were also characterized by spa-types. Minimum inhibitory concentrations (MICs) to chlorhexidine were determined by the broth dilution method. Of 520 isolates collected, 6·2% were methicillin-resistant S. aureus (MRSA, all mecA-positive) and mupirocin resistance was low [0·8%, 95% confidence interval (CI) 0·3–2·0] and only found in MRSA. Carriage of the qacA gene was identified in 1·7% (95% CI 0·8–3·3) of isolates and 3·5% (95% CI 2·2–5·4) had a chlorhexidine MIC of 4 mg/l. The PVL gene was infrequent (3·7%, 95% CI 2·4–5·6). Genotyping identified 234 spa-types that mapped to 22 clonal complexes. Comparison of these current data with previous work suggest that the widespread use of staphylococcal decolonization regimens over the past decade or more has not had an adverse impact on resistance rates, PVL carriage or the prevalence of specific S. aureus lineages.


Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 430
Author(s):  
Wichai Santimaleeworagun ◽  
Praewdow Preechachuawong ◽  
Wandee Samret ◽  
Tossawan Jitwasinkul

Methicillin-resistant Staphylococcus aureus (MRSA) is mostly found in Thailand in the hospital as a nosocomial pathogen. This study aimed to report the genetic characterization of a clinical community-acquired MRSA (CA-MRSA) isolate collected from hospitalized patients in Thailand. Among 26 MRSA isolates, S. aureus no. S17 preliminarily displayed the presence of a staphylococcal cassette chromosome mec (SCCmec) type IV pattern. The bacterial genomic DNA was subjected to whole-genome sequencing. Panton–Valentine leukocidin (PVL) production, virulence toxins, and antibiotic resistance genes were identified, and multi-locus sequence typing (MLST) and spa typing were performed. The strain was matched by sequence to MLST type 2885 and spa type t13880. This strain carried type IV SCCmec with no PVL production. Five acquired antimicrobial resistance genes, namely blaZ, mecA, Inu(A), tet(K), and dfrG conferring resistance to β-lactams, lincosamides, tetracycline, and trimethoprim, were identified. The detected toxins were exfoliative toxin A, gamma-hemolysin, leukocidin D, and leukocidin E. Moreover, there were differences in seven regions in CR-MRSA no. S17 compared to CA-MRSA type 300. In summary, we have reported the ST2885-SCCmec IV CA-MRSA clinical strain in Thailand for the first time, highlighting the problem of methicillin resistance in community settings and the consideration in choosing appropriate antibiotic therapy.


2000 ◽  
Vol 44 (2) ◽  
pp. 294-303 ◽  
Author(s):  
Richard F. Pfeltz ◽  
Vineet K. Singh ◽  
Jennifer L. Schmidt ◽  
Michael A. Batten ◽  
Christopher S. Baranyk ◽  
...  

ABSTRACT A series of 12 Staphylococcus aureus strains of various genetic backgrounds, methicillin resistance levels, and autolytic activities were subjected to selection for the glycopeptide-intermediate S. aureus (GISA) susceptibility phenotype on increasing concentrations of vancomycin. Six strains acquired the phenotype rapidly, two did so slowly, and four failed to do so. The vancomycin MICs for the GISA strains ranged from 4 to 16 μg/ml, were stable to 20 nonselective passages, and expressed resistance homogeneously. Neither ease of acquisition of the GISA phenotype nor the MIC attained correlated with methicillin resistance hetero- versus homogeneity or autolytic deficiency or sufficiency. Oxacillin MICs were generally unchanged between parent and GISA strains, although the mec members of both isogenic methicillin-susceptible and methicillin-resistant pairs acquired the GISA phenotype more rapidly and to higher MICs than did their susceptible counterparts. Transmission electron microscopy revealed that the GISA strains appeared normal in the absence of vancomycin but had thickened and diffuse cell walls when grown with vancomycin at one-half the MIC. Common features among GISAs were reduced doubling times, decreased lysostaphin susceptibilities, and reduced whole-cell and zymographic autolytic activities in the absence of vancomycin. This, with surface hydrophobicity differences, indicated that even in the absence of vancomycin the GISA cell walls differed from those of the parents. Autolytic activities were further reduced by the inclusion of vancomycin in whole-cell and zymographic studies. The six least vancomycin-susceptible GISA strains exhibited an increased capacity to remove vancomycin from the medium versus their parent lines. This study suggests that while some elements of the GISA phenotype are strain specific, many are common to the phenotype although their expression is influenced by genetic background. GISA strains with similar glycopeptide MICs may express individual components of the phenotype to different extents.


2021 ◽  
Author(s):  
Jayshree Singh ◽  
Amit Kumar ◽  
Sharad K. Yadav ◽  
Ritika Yadav ◽  
Vinod K. Singh

Abstract Staphylococcus aureus has been described as the most common cause of human and animal diseases and has emerged as superbug due to multidrug resistance. Considering these, a total of 175 samples were collected from pyogenic cases of humans (75) and animals (100), to establish the drug resistance pattern and also for molecular characterization of human and animal isolates. Thermonuclease (nuc) gene amplification was used to confirm all presumptive S. aureus isolates and then antibiotic sensitivity and slide coagulase tests were used for phenotypic characterization of isolates. Following that, all of the isolates were subjected to PCR amplification to detect the existence of the methicillin resistance (mecA) and coagulase (coa) genes. Lastly, typing was done by using the Randomly Amplified Polymorphic DNA-PCR. The overall prevalence of S. aureus in human and animal samples was found to be 39.4%. Drug sensitivity revealed the highest resistance against the β-lactam antibiotics such as ampicillin (94.8%) and penicillin (90.6%), followed by cephalosporin (cefixime-67.7%) and quinolone (ciprofloxacin-52.1%) group of drugs. The drug sensitivity was the highest against antibiotics like chloramphenicol (95%) followed by gentamicin (90%). Among the 69 S. aureus isolates, the overall presence of MRSA was 40.5% (27.5% and 50% in human and animal isolates respectively). Total 33 isolates exhibited coa genes amplification of more than one amplicons and variable in size of 250, 450, 800, and 1100 bp. The RAPD typing revealed amplification of 5 and 6 different band patterns in humans and animals, respectively, with two common patterns suggesting a common phylogenetic profile.


2008 ◽  
Vol 190 (18) ◽  
pp. 6188-6196 ◽  
Author(s):  
Ian R. Siboo ◽  
Donald O. Chaffin ◽  
Craig E. Rubens ◽  
Paul M. Sullam

ABSTRACT The SraP adhesin of Staphylococcus aureus is a member of a highly conserved family of serine-rich surface glycoproteins of gram-positive bacteria. For streptococci, export of the SraP homologs requires a specialized transport pathway (the accessory Sec system). Compared to streptococci, however, SraP is predicted to differ in its signal peptide and glycosylation, which may affect its dependence on a specialized system for transport. In addition, two genes (asp4 and asp5) essential for export in Streptococcus gordonii are missing in S. aureus. Thus, the selectivity of the accessory Sec system in S. aureus may also differ compared to streptococci. To address these issues, the five genes encoding the putative accessory Sec system (secY2, secA2, and asp1-3) were disrupted individually in S. aureus ISP479C, and the resultant mutants were examined for SraP export. Disruption of secA2 resulted in the near complete loss of SraP surface expression. Similar results were seen with disruption of secY2 and asp1, asp2, or asp3. To assess whether the accessory Sec system transported other substrates, we compared secreted proteomes of ISP479C and a secA2 isogenic mutant, by two-dimensional fluorescence difference gel electrophoresis. Although two consistent differences in proteome content were noted between the strains, neither protein appeared to be a likely substrate for accessory Sec export. Thus, the accessory Sec system of S. aureus is required for the export of SraP, and it appears to be dedicated to the transport of this substrate exclusively.


2008 ◽  
Vol 19 (3) ◽  
pp. 227-232 ◽  
Author(s):  
Angela M Sloan ◽  
Averil M Henderson ◽  
Raymond SW Tsang

INTRODUCTION: Serogroup ANeisseria meningitidishas repeatedly caused epidemics of invasive meningococcal disease (IMD) in developing nations since the 1960s. The present study is the first detailed study of serogroup A bacteria isolated in Canada.METHODS: Thirty-four serogroup A meningococcal isolates collected from individuals with IMD in Canada between 1979 and 2006 were characterized by serology and multilocus sequence typing of seven housekeeping enzyme genes and genes encoding three outer membrane protein antigens.RESULTS: Isolates were assigned to either the sequence type (ST)-1 or the ST-5 clonal complex. Clones within the ST-1 complex were recovered between 1979 and 1992, while clones of the ST-5 complex were isolated between 1987 and 2006; respectively, they accounted for 70.6% and 29.4% of all isolates studied. Isolates of the ST-1 complex were characterized by serosubtype antigen P1.3 or P1.3,6 with PorB allele 60 (serotype 4) and FetA sequence F5-1, while isolates of the ST-5 complex were characterized by serosubtype antigen P1.9 with PorB allele 47 (also serotype 4) and FetA sequence F3-1.CONCLUSIONS: The Canadian serogroup A IMD isolates likely originated in travellers returning from hyperendemic or epidemic areas of the globe where serogroup A bacteria circulate. Although the Canadian cases of serogroup A IMD were caused by clones known to have caused epidemics in developing countries, disease incidence remained low in Canada.


2015 ◽  
Vol 6 ◽  
Author(s):  
Adebayo O. Shittu ◽  
Omotayo Oyedara ◽  
Kenneth Okon ◽  
Adeola Raji ◽  
Georg Peters ◽  
...  

2017 ◽  
Vol 36 (10) ◽  
pp. 1787-1793 ◽  
Author(s):  
C. L. Hanis ◽  
K. E. Garrett ◽  
H. T. Essigmann ◽  
D. A. Robinson ◽  
S. M. Gunter ◽  
...  

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