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ABSTRACTTyping of methicillin-resistant Staphylococcus aureus (MRSA) is important in infection control and surveillance. The current nomenclature of MRSA includes the genetic background of the S. aureus strain determined by multilocus sequence typing (MLST) or equivalent methods like spa typing and typing of the mobile genetic element staphylococcal cassette chromosome mec (SCCmec), which carries the mecA or mecC gene. Whereas MLST and spa typing are relatively simple, typing of SCCmec is less trivial because of its heterogeneity. Whole-genome sequencing (WGS) provides the essential data for typing of the genetic background and SCCmec, but so far, no bioinformatic tools for SCCmec typing have been available. Here, we report the development and evaluation of SCCmecFinder for characterization of the SCCmec element from S. aureus WGS data. SCCmecFinder is able to identify all SCCmec element types, designated I to XIII, with subtyping of SCCmec types IV (2B) and V (5C2). SCCmec elements are characterized by two different gene prediction approaches to achieve correct annotation, a Basic Local Alignment Search Tool (BLAST)-based approach and a k-mer-based approach. Evaluation of SCCmecFinder by using a diverse collection of clinical isolates (n = 93) showed a high typeability level of 96.7%, which increased to 98.9% upon modification of the default settings. In conclusion, SCCmecFinder can be an alternative to more laborious SCCmec typing methods and is freely available at https://cge.cbs.dtu.dk/services/SCCmecFinder.IMPORTANCE SCCmec in MRSA is acknowledged to be of importance not only because it contains the mecA or mecC gene but also for staphylococcal adaptation to different environments, e.g., in hospitals, the community, and livestock. Typing of SCCmec by PCR techniques has, because of its heterogeneity, been challenging, and whole-genome sequencing has only partially solved this since no good bioinformatic tools have been available. In this article, we describe the development of a new bioinformatic tool, SCCmecFinder, that includes most of the needs for infection control professionals and researchers regarding the interpretation of SCCmec elements. The software detects all of the SCCmec elements accepted by the International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements, and users will be prompted if diverging and potential new elements are uploaded. Furthermore, SCCmecFinder will be curated and updated as new elements are found and it is easy to use and freely accessible.

Modern cloud computing environments require strong cryptographic solutions to ensure data confidentiality and security against quantum computing threats. This work proposes a new cryptographic framework that combines Supersingular Isogeny Diffie–Hellman (SIDH) with certificateless hashed public key authenticated encryptions with keyword search to improve the need of security and efficiency in cloud computing environments. The majority of conventional cryptographic solutions have vulnerability under quantum computing attacks and inefficiency in key management and keyword search. This framework has exploited SIDH to overcome the drawbacks of the approaches. By exploiting the inherent computational difficulties in isogenies between supersingular elliptic curves, strong post-quantum security is gained. It assures the system against the classical and quantum threats. Searching on encrypted data using SIDH with CL-HPAEKS is more secure and efficient. CL-HPAEKS aims at providing a certificate-less secure keyword search system to eliminate the overhead associated with certificate management while keeping high security standards. The result shows an average time to be around 534–4334 ms for encryption and from 812 to 4041 ms for decryption, depending on the size of data.

Owls are nocturnal predators that inhabit urbanized and farmlands. They are in direct contact with other animals, both livestock and small wild rodents that they mostly feed on. Staphylococci can be both commensal and pathogenic bacteria that are widespread across the various ecological niches. We aimed to isolate staphylococci from owls and to characterize their antimicrobial resistance, virulence factors and genetic lineages. Swab samples were collected from the throat and cloaca of 114 owls admitted to two rehabilitation centers in Portugal. The identification of staphylococci species was performed by MALDI-TOF. Staphylococci antimicrobial resistance and virulence genes were investigated by means of the disk diffusion method and PCR. Staphylococcus aureus isolates were characterized by MLST, agr and spa-typing. Of the tested animals, 66 isolates were recovered, including 10 different species of staphylococci, of which 25 were coagulase-positive (CoPS) and 41 were coagulase-negative (CoNS). Twenty-three S. aureus were isolated, of which one mecC-MRSA was identified. The isolates were mainly resistant to penicillin, aminoglycosides, clindamycin and tetracycline. mecC-MRSA belonged to ST1245 and spa-type t843 and the remaining S. aureus were ascribed to 12 STs and 15 spa types. A high diversity of clonal lineages was identified among the S. aureus isolated from wild owls. Owls feed mainly on small rodents often exposed to waste and anthropogenic sources, which may explain the moderate prevalence of S. aureus in these animals.

Background: The availability of comprehensive data on the ecology and molecular epidemiology of Staphylococcus aureus/MRSA in wild animals is necessary to understand their relevance in the “One Health” domain. Objective: In this study, we determined the pooled prevalence of nasal, tracheal and/or oral (NTO) Staphylococcus aureus (S. aureus) and methicillin-resistant S. aureus (MRSA) carriage in wild animals, with a special focus on mecA and mecC genes as well as the frequency of MRSA and methicillin susceptible S. aureus (MSSA) of the lineages CC398 and CC130 in wild animals. Methodology: This systematic review was executed on cross-sectional studies that reported S. aureus and MRSA in the NTO cavities of wild animals distributed in four groups: non-human primates (NHP), wild mammals (WM, excluding rodents and NHP), wild birds (WB) and wild rodents (WR). Appropriate and eligible articles published (in English) between 1 January 2011 to 30 August 2021 were searched for from PubMed, Scopus, Google Scholar, SciElo and Web of Science. Results: Of the 33 eligible and analysed studies, the pooled prevalence of NTO S. aureus and MRSA carriage was 18.5% (range: 0–100%) and 2.1% (range: 0.0–63.9%), respectively. The pooled prevalence of S. aureus/MRSA in WM, NHP, WB and WR groups was 15.8/1.6, 32.9/2.0, 10.3/3.4 and 24.2/3.4%, respectively. The prevalence of mecC-MRSA among WM/NHP/WB/WR was 1.64/0.0/2.1/0.59%, respectively, representing 89.9/0.0/59.1/25.0% of total MRSA detected in these groups of animals.The MRSA-CC398 and MRSA-CC130 lineages were most prevalent in wild birds (0.64 and 2.07%, respectively); none of these lineages were reported in NHP studies. The MRSA-CC398 (mainly of spa-type t011, 53%), MRSA-CC130 (mainly of spa types t843 and t1535, 73%), MSSA-CC398 (spa-types t571, t1451, t6606 and t034) and MSSA-CC130 (spa types t843, t1535, t3625 and t3256) lineages were mostly reported. Conclusion: Although the global prevalence of MRSA is low in wild animals, mecC-mediated resistance was particularly prevalent among MRSA isolates, especially among WM and WB. Considering the genetic diversity of MRSA in wild animals, they need to be monitored for effective control of the spread of antimicrobial resistance.

Methicillin-resistant Staphylococcus aureus (MRSA) is a clinically prevalent bacterium and is resistant to many drugs. Genetic factors such as mec genes are considered to be responsible for this resistance. Recently, Staphylococcal Cassette Chromosome mec (SCCmec) element mutations produced mecC, a new genetic variant that encodes a transpeptidase enzyme (63% similarity with mecA-encoded PBP2a). This cross-sectional study was conducted to establish the prevalence of the mecA and mecC genes among phenotypically identified MRSA and their effectiveness against different antibiotics in clinical specimens. The prevalence of Staphylococcus aureus was 10.2% (n = 102) in the total number of clinical specimens collected (n = 1000). However, the prevalence of MRSA was 6.3% (n = 63) of the total samples collected, while it was 61.8% among total Staphylococcus aureus isolates. mec genes were confirmed in 96.8% (n = 61) isolates of MRSA, while 3.2% (n = 2) were found to be negative for mec genes. The combination of mecA and mecC was detected in 57.1% (n = 36) of the MRSA isolates. The prevalence of lone mecA was 31.8% (n = 20) and that of lone mecC was 7.9% (n = 5) among all the MRSA samples. Penicillin and amoxicillin/clavulanic acid were the most resistant antibiotics followed by norfloxacin (91.2%), levofloxacin (87.1%), ciprofloxacin (83.9%), azithromycin (78.6%), erythromycin (77.4%), moxifloxacin (69.8%), and sulfamethoxazole/trimethoprim (54.9%). On the other hand, vancomycin and teicoplanin (98.4%) were more effective drugs against MRSA followed by linezolid (96.7%), clindamycin (84.6%), chloramphenicol (83.7%), fusidic acid (70.6%), gentamicin (67.7%), and tetracycline (56.8%). In conclusion, a significant prevalence of mecA and mecC has been found among MRSA isolated from clinical specimens, which is likely responsible for antibiotic resistance in MRSA in our clinical settings. However, vancomycin, teicoplanin, and linezolid were found the top three most effective drugs against MRSA in our clinical settings. Thus, MRSA endemics in local areas require routine molecular and epidemiological investigation.

Several methicillin‐resistant Staphylococcus aureus (MRSA) lineages that carry a novel mecA homologue ( mecC ) have recently been described in livestock and humans. In Denmark, two independent human cases of mecC ‐MRSA infection have been linked to a livestock reservoir. We investigated the molecular epidemiology of the associated MRSA isolates using whole genome sequencing (WGS). Single nucleotide polymorphisms (SNP) were defined and compared to a reference genome to place the isolates into a phylogenetic context. Phylogenetic analysis revealed two distinct farm‐specific clusters comprising isolates from the human case and their own livestock, whereas human and animal isolates from the same farm only differed by a small number of SNPs, which supports the likelihood of zoonotic transmission. Further analyses identified a number of genes and mutations that may be associated with host interaction and virulence. This study demonstrates that mecC ‐MRSA ST130 isolates are capable of transmission between animals and humans, and underscores the potential of WGS in epidemiological investigations and source tracking of bacterial infections. →See accompanying article http://dx.doi.org/10.1002/emmm.201302622 Graphical Abstract The promise of whole genome sequencing in epidemiological investigations and the source tracking of bacterial infections is exemplified here by the discovery of potential zoonotic transmission of methicillin‐resistant Staphylococcus aureus strains.

The aim of this study was to determine the carriage rate of coagulase-positive staphylococci (CoPS) in wild birds and to characterize recovered isolates. Tracheal samples from 324 wild birds, obtained in different Spanish regions during 2015–2016, were screened for CoPS carriage. The antimicrobial resistance profile and the virulence gene content were investigated. Molecular typing was performed by spa, agr, MLST, SCCmec, and S. delphini group classification. CoPS were recovered from 26 samples of wild birds (8.3%), and 27 isolates were further characterized. Two CoPS species were detected: S. aureus (n = 15; eight cinereous vultures and seven magpies) and S. delphini (n = 12; 11 cinereous vultures and one red kite). Thirteen S. aureus were methicillin-resistant (MRSA) and the remaining two strains were methicillin-susceptible (MSSA). Twelve MRSA were mecC-positive, typed as t843-ST1583/ST1945/ST1581/ST1571 (n = 11) and t1535-ST1945 (n = 1) (all of clonal-complex CC130); they were susceptible to the non-ß-lactams tested. The remaining MRSA strain carried the mecA gene, was typed as t011-ST398-CC398-agrI-SCCmec-V, and showed a multiresistance phenotype. MSSA isolates were ascribed to lineages ST97-CC97 and ST425-CC425. All S. aureus lacked the studied virulence genes (lukS/F-PV, tst, eta, etb, and etd), and the IEC type E (with scn and sak genes) was detected in four mecC-positive and one MSSA isolates. S. delphini strains were methicillinsusceptible but showed resistance to at least one of the antimicrobials tested, with high penicillin (75%, with blaZ gene) and tetracycline [58%, with tet (K)± tet (L)] resistance rates. All S. delphini isolates presented the virulence genes lukS-I, siet, and seint, and four carried the clindamycin-resistance lnu (A) gene.

Livestock associated Methicillin resistant Staphylococcus aureus (S. aureus) (LA-MRSA) was reported to be zoonotic and may transmit to farmers and veterinarians. The objectives of this study were to investigate the occurrence of LA-MRSA from dairy cattle and to evaluate the antimicrobial resistance profiles of the isolates. A total of 63 milk and 32 nasal swab samples were randomly collected from dairy cattle. The samples were processed to isolate S. aureus, MRSA and LA-MRSA using both phenotypic and molecular methods using PCR. The confirmed S. aureus isolates were cultured on oxacillin resistant screening agar base (ORSAB) to detect MRSA and the isolates were further confirmed by PCR targeting the mecA gene. Detection of the novel mecA gene, mecC gene was conducted by PCR amplification. The antimicrobial susceptibility tests were conducted using disc diffusion method. Results revealed 17/95 (17.89%) and 15/95 (15.79%) were positive for mecA and mecC genes respectively. Out of the 15 mecC positive isolates, 12 were positive for both mecA and mecC. The MRSA isolates showed multidrug resistance. The findings showed high prevalence of mecC-positive LA-MRSA in Malaysia and highlight the public health risks to people that may come in contact with the carrier animals or those who may consume unpasteurized milk products from these animals.

Background Staphylococcus aureus strains are highly virulent and associated with an eclectic range of severe nosocomial and community-acquired infections. Objectives This study assessed methicillin- and vancomycin-resistant Staphylococcus aureus (MRSA/VRSA) from clinical and ready-to-eat (RTE) food sources, screened for antibiotic resistance; and molecular determinants of antibiotic and virulence genes. Methods Altogether, 465 clinical and RTE food samples were analyzed via conventional microbiological techniques and S. aureus identification was confirmed by nuc gene detection. Phenotypic screening for methicillin and vancomycin-resistance was by agar-screen cum micro-broth dilution respectively, while antibiotic susceptibility testing was done by the disc-diffusion technique. VanA/vanB/VanC1 , femA , mecA/mecC; pvl/hlg and spa gene detection was via Polymerase chain reaction. Results Phenotypically, 211 Staphylococcal isolates were recovered, 138 (65.4%) of them carrying the nuc gene – all 138 (100.0%) were VRSA, while 59/138 (42.8%) were MRSA phenotypically. Overall, 114/138 (82.6%), 7/138 (5.1%), and 6/138 (4.3%) of isolates had the femA , mecA , and mecC genes, while van genes were detected in only 3 (2.2%) isolates, with virulence determinants pvl , hlg , and spa gene carriage in 8 (5.8%), 10 (7.2%), and 77 (55.8%) isolates respectively. In all, 11.6% carried resistance-associated genes, 55.8% carried virulence genes, and co-detection of resistance and virulence genes was observed in 12.3%. Overall, 96/138 (69.6%) were multidrug-resistant (MDR), while one strain was extremely drug-resistant (XDR). MAR Indices ≥ 0.2 was observed in 83.3% of isolates. Conclusion This study highlights virulence levels of MRSA and VRSA circulating strains in Osogbo, contributing to their sustained surveillance, and improving available data for successive epidemiology investigations. This study also reports the occurrence of the mecC gene in S. aureus isolates from RTE foods and human samples in Southwestern Nigeria.