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As with IoT devices, cloud computation increases its applicability to other areas that use the information and puts it in a commonplace that is necessary for computing and analysis. These devices need the cloud to store and retrieve data since they are unable to store and process data on their own. Cloud computing offers a variety of services to consumers, including IaaS, PaaS, and SaaS. The usage of cloud resources for data storage that may be accessible by all users associated with cloud computing is a key disadvantage of cloud computing. Without disclosing the data's contents, the usage of Public Key Encryptions with Keyword Search (PEKS) secures publicly encrypted keys against third-party search capabilities that are not trustworthy. PEKs provide a security risk every time Interior Keywords Guessing Attacks (IKGA) are discovered in the system since an unauthorized service estimates the keywords in the trapdoor. This issue can be resolved using various methodologies such as the Certificateless Fleshed Public Key Authenticated Encryption of Keyword Search (CL-HPAEKS), which also uses Altered Elliptic Curve Cryptography (MECC), and the Mutation Centred Flower Pollinations Algorithm (CM-FPA), which uses optimization in keys to be improving the algorithm's performance. The system's security is achieved by adding a Message Fragments 5 (MD5) scramble mechanism. The proposed system achieves system security of 96%, and it takes less time to implement than earlier encryption methods. [ABSTRACT FROM AUTHOR]

The aim of the present study was to determine the genetic relatedness between Methicillin-resistant Staphylococcus aureus (MRSA) isolated from pet animals and human. In that context, 561 swabs were collected from dogs (n=238) and cats (n=323). In addition, 30 MRSA positive isolates from Qasr Elainy hospital, from different patients were also included. The collected samples were subjected for bacteriological examination for isolation of S. aureus. The recovered S. aureus isolates that were resistant to oxacillin and cefoxitin were tested for the presence of mecA, mecC, spa, and nuc genes by PCR, followed by a sequence analysis of the mecA gene in seven selected isolates. Out of 238 swab samples collected from dogs and 323 swab samples collected from cats, 13 (5.46%) and 8 (2.48%) isolates respectively were confirmed to be MRSA. In addition, the 30 human isolates were all categorized as MRSA. The mecA gene was successfully identified in all isolates from dogs, cats and human, while spa was harbored by 13, 4 and 30 isolates from dogs, cats and human respectively. Furthermore 13, 4 and 23 isolates from dogs, cats and human were positive for the nuc gene. mecC was not identified from all examined isolates. The phylogenetic analysis of the mecA gene sequence in seven selected MRSA isolates demonstrated a high degree of similarity among isolates originating from dogs, cats, and humans. This study confirms the zoonotic spread of S. aureus and suggests that dogs and cats are probably contributing to the spread of MRSA to human. [ABSTRACT FROM AUTHOR]

Methicillin-resistant Staphylococcus aureus (MRSA) is a frequent cause of healthcare- and community-associated infections. Nasal carriage of MRSA is a risk factor for subsequent MRSA infections. Increased morbidity and mortality are associated with MRSA infections and screening and diagnostic tests for MRSA play an important role in clinical management. A literature search was conducted in PubMed and supplemented by citation searching. In this article, we provide a comprehensive review of molecular-based methods for MRSA screening and diagnostic tests including individual nucleic acid detection assays, syndromic panels, and sequencing technologies with a focus on their analytical performance. Molecular based-assays for the detection of MRSA have improved in terms of accuracy and availability. Rapid turnaround enables earlier contact isolation and decolonization for MRSA. The availability of syndromic panel tests that include MRSA as a target has expanded from positive blood cultures to pneumonia and osteoarticular infections. Sequencing technologies allow detailed characterizations of novel methicillin-resistance mechanisms that can be incorporated into future assays. Next generation sequencing is capable of diagnosing MRSA infections that cannot be identified by conventional methods and metagenomic next-generation sequencing (mNGS) assays will likely move closer to implementation as front-line diagnostics in the near future. [ABSTRACT FROM AUTHOR]

ABSTRACT: Objectives: In this study, we aimed to assess the extent of dissemination of methicillin-resistant Mammaliicoccus sciuri in animal farms in Tunisia and evaluate the distribution of virulence and methicillin resistance genes in the M. sciuri population. Methods: Staphylococci and mammaliicocci isolated from unhealthy animals and healthy humans from adjacent farms in Tunisia were characterized for antimicrobial susceptibility, biofilm formation, agglutination, and hemolysis abilities. Mammaliicoccus sciuri relatedness and content in antibiotic resistance and virulence genes were analyzed by whole-genome sequencing (WGS). Results: Mammaliicoccus sciuri was the most prevalent species (46.2%), showing the highest resistance rates to fusidic acid (94.6%), oxacillin (73%), penicillin (40.5%), clindamycin (37%), ciprofloxacin (27%), and cefoxitin (24.3%). Some isolates carried genes encoding resistance to nine different antibiotic classes. mecA was found in 35% of M. sciuri and mecC in 16.2%. All isolates carrying mecC were of S. sciuri subspecies carnaticus and carried the hybrid element SCCmec-mecC. Mammaliicoccus sciuri were able to produce strong biofilm (27%) and have clumping ability (16%). Additionally, they carried genes for capsule production (cap8, 100%), iron-regulated surface determinants (isdE, 24%; isdG, 3%), and virulence regulation (clpC and clpP, 100%). Single nucleotide polymorphisms (SNPs) analysis showed that 17 M. sciuri cross-transmission events probably occurred between different animal species and farms. Moreover, SCCmec was estimated to have been acquired five times by S. sciuri subsp. carnaticus. Conclusion: Multidrug resistant and pathogenic M. sciuri were frequently disseminated between different animal species within the farm environment. mecA and mecC can be disseminated by both frequent acquisition of the SCCmec element and clonal dissemination.

This study was conducted for the molecular detection of the mecA, mecC, and nuc gene among MRSA and to investigate biofilm formation among the methicillin-resistant Staphylococcus aureus (MRSA) clinical isolates. A total of 208 different samples were collected and processed for phenotypic and genotypic identification of MRSA. All MRSA isolates were subjected to antibiotics sensitivity, cefoxitin disk diffusion test, and vancomycin minimum inhibitory concentration (MIC) E-test. The MRSA isolates were detected for the presence of mecA, mecC, and nuc genes. Congo red agar (CRA) method was used to assess the ability of isolates to form biofilms. The results of the study showed that the prevalence of MRSA was 48%. The MRSA isolates were highly resistant (100%) to penicillin, ß lactamase inhibitors, cephalosporins, and macrolides. All the MRSA isolates were susceptible to vancomycin antibiotic drugs. Cefoxitin (30 µg) disk diffusion test showed 100% sensitivity and specificity for the identification of MRSA phenotypically. A total of 100 MRSA clinical isolates were positive for the mecA and nuc gene. Only 3 MRSA isolates were positive for the mecC gene. Congo red agar method showed that 20 (20%) isolates formed moderate biofilm while 80 (80%) isolates were non-biofilm forming. Multi drugs resistant and mecC gene-positive MRSA isolates are rapidly emerging in Pakistan. Therefore, the mecC gene should be detected along with the mecA gene for the identification of MRSA clinical isolates. It also requires early identification of biofilm formation and necessary interventions for its effective treatment and control. [ABSTRACT FROM AUTHOR]

Methicillin-resistant in Staphylococci is a serious public health issue. It is mostly encoded by the mecA gene. The mecC gene is a new mecA analog responsible for resistance to methicillin in some Staphylococcal clinical isolates. This mecC gene is still underestimated in Egypt. The aim of the current study was to detect mecA and mecC genes in clinical Staphylococci isolates from a tertiary care university hospital in Egypt compared to the different phenotypic methods. A total of 118 Staphylococcus aureus (S. aureus) and 43 coagulase-negative Staphylococci (CoNS) were identified from various hospital-acquired infections. Methicillin resistance was identified genotypically using the PCR technique and phenotypically using the cefoxitin disc diffusion test, oxacillin broth microdilution and the VITEK2 system in all Staphylococcal isolates. The mecA gene was detected in 82.2% of S. aureus and 95.3% of CoNS isolates, while all of the isolates tested negative for the mecC gene. Interestingly, 30.2% of CoNS isolates showed the unique character of inducible oxacillin resistance, being mecA-positive but oxacillin-susceptible (OS-CoNS). The dual use of genotypic and phenotypic methods is highly recommended to avoid missing any genetically divergent strains. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Methicillin-resistant Staphylococcus aureus bearing the mecC gene (mecC-MRSA) has been reported from animals and humans in recent years. This study describes the first mecC-MRSA isolates of human and equine origin in Hungary (two isolates from horses and one from a veterinarian, who treated one of the infected horses, but was asymptomatic). MRSA isolates were identified by cultivation and PCR detection of the species-specific spa gene and mecA/mecC methicillin resistance genes. The isolates were characterized by antibiotic susceptibility testing, MLST, spa, SCCmec typing, PFGE and whole genome sequencing (WGS). All three isolates belonged to the ST130-t843-SCCmec XI genotype, and carried the mecC and blaZ genes. Apart from beta-lactam drugs, they were sensitive to all tested antibiotics. The isolates of the infected horse and its veterinarian had the same PFGE pulsotype and showed only slight differences with WGS. Hence, this is the first description of direct transmission of a mecC-carrying MRSA between a horse and its veterinarian. The emergence of mecC in the country highlights the importance of the appropriate diagnostics in MRSA identification. [ABSTRACT FROM AUTHOR]

Objective: Compare the use of blood products and intravenous fluid management in patients scheduled for coronary artery bypass surgery and randomized to minimal invasive extracorporeal circulation (MiECC) and conventional extracorporeal circulation (CECC). Methods: A total of 240 patients who were scheduled for their first on-pump CABG, were randomized to MiECC or CECC groups. The study period was the first 84 hours after surgery. Hemoglobin <80 g/l was used as transfusion trigger. Results: Red blood cell transfusions intraoperatively were given less often in the MiECC group (23.3% vs 9.2%, p = 0.005) and the total intravenous fluid intake was significantly lower in the MiECC group (3300 ml [2950–4000] vs 4800 ml [4000–5500], p < 0.001). Hemoglobin drop also was lower in the MiECC group (35.5 ± 8.9 g/l vs 50.7 ± 9 g/l, p < 0.001) as was hemoglobin drop percent (25.3 ± 6% vs 35.3 ± 5.9%, p < 0.001). Chest tube drainage output was higher in the MiECC group (645 ml [500–917.5] vs 550 ml [412.5–750], p = 0.001). Particularly, chest tube drainage in up to 600 ml category, was in benefit of CECC group (59.1% vs 40.8%, p = 0.003). ROC curve analysis showed that patients with hemoglobin level below 95 g/l upon arrival to intensive care unit was associated with increased risk of developing postoperative atrial fibrillation (POAF) (p = 0.002, auc = 0.61, cutoff <95, sensitivity = 0.47, positive predictive value = 0.64). Conclusion: MiECC reduced the intraoperative need for RBC transfusion and intravenous fluids compared to the CECC group, also reducing hemoglobin drop compared to the CECC group in CABG surgery patients. Postoperative hemoglobin drop was a predictor of POAF. [ABSTRACT FROM AUTHOR]

The risk of mortality and morbidity is high in patients with advanced age and additional systemic diseases who will undergo coronary artery bypass surgery. For this reason, the chosen anesthesia method and surgical approach gain importance. With the “Fast track cardiac anesthesia” (FTKA) protocol, the length of stay in hospitals and intensive care units is shortened, thus reducing the risk of mortality and morbidity in elderly and comorbid patients. In recent years, the use of Minimal Extracorporeal Circulatory Systems (MECC) has become widespread by modifying conventional extracorporeal circulatory systems in open heart surgery. With MECC, the systemic inflammatory response seen in the conventional system is reduced. In this article, we present the combination of FTCA protocol and MECC use, which we successfully applied in an 81-year-old patient with comorbidities. Our patient underwent triple coronary artery bypass surgery and was discharged on the 6th post-operative day with full recovery. We believe that the risks can be reduced and successful results are obtained with the anesthesia method (FTKA) and surgical approach (MECC) chosen in patients with advanced age and additional systemic diseases who will undergo coronary artery bypass surgery. [ABSTRACT FROM AUTHOR]

Objective:The diagnosis and treatment of mecC positive methicillin resistant Staphylococcus aureus (MRSA) isolates pose a significant problem in clinical microbiology and infectious disease practices. The studies on the frequency of mecC positive isolates in Turkey is rather scarce. In the present study, we aimed to investigate the presence of mecA, mecC, spa and pvu genes in MRSA strains isolated from various clinical specimens submitted to Clinical Microbiology Laboratories of Bezmialem Vakıf Hospital.Methods:We performed nucleic acid extraction and multiplex polymerase chain reaction (PCR) to 126 MRSA strains to detect mecC, mecA, spa and pvl genes.Results:According to the multiplex PCR results of 126 MRSA strains studied, 126 (100%) had mecA, 107 (85%) had spa, and 3 (2%) had pvl genes. We performed another polymerase chain reaction protocol and spa genes were identified in 19 of specimens, which were found negative priorly.Conclusion:Considering the factors that a university medical center where the study was conducted provided a tertiary healthcare service to a large metropolitan area in Istanbul and none of the isolates carried mecC gene might indicate that mecC gene carrying MRSA isolates did not pose a significant public health threat in Turkey.

Nowadays, bovine mastitis has been recorded as an inflammation of the mammary gland and Staphylococcus aureus is the main pathogens responsible for contagious mastitis in ruminants. Presented study is looking for to assess the existence of methicillin-resistant and PVL positive Staphylococcus aureus from bovine milk and if there is any relation among the detected genes and antibiotic resistance or not? For this purpose, 100 milk samples were collected from cows affected with mastitis from four dairy farms in Fars province. The samples were transferred to MSA medium and incubated at 37 °C for 24 h. The pure colonies were identified by biochemical as well as molecular techniques using mecA, mecC and PVL gene. Then, the antibiotic pattern of the isolates was analyzed based on CLSI 2019 and Whonet program. Finally, the results were analyzed using statistical analysis. Totally, 6 S. aureus strains were detected which was confirmed by biochemical as well as molecular techniques. The isolates showed different antibiotic susceptibility patterns, most of the isolates were multidrug resistant, while, the strain No. 5 was the most sensitive one. Our results indicated presence of mecA gene in strains 1, 2 and 3, mecC in isolates 4, 5 and 6, and the PVL gene only in strain 4. Hence, it could be concluded that although different phages transporting PVL genes to the S. aureus isolate, in this special research we didn’t find any relationship between presence of mecA, mecC, and PVL genes with antibiotics resistant pattern.

Coagulase positive Staphylococci (CoPS) are the leading cause of canine cutaneous and otic infections. Virulence factors associated with Staphylococci include the expression of mec and panton-valentine leukocidin (pvl) genes. Methicillin-resistance (MR) is commonly associated with mecA gene expression, although a recently identified variant, mecC, has been reported. This study aims to evaluate the prevalence of mecA, mecC and pvl genes in 232 clinical isolates of CoPS collected from dogs with pyoderma. A multiplex PCR, and Kirby-Bauer disk diffusion susceptibility test for cefoxitin was performed for all isolates. PBP2a agglutination test was performed on 127 isolates. Standard MRSA isolates were used as positive controls. The mecA gene was identified in 149/232 isolates (64.2%): 116 S. pseudintermedius, 30 S. coagulans and three S. aureus. The pvl gene was present in only 1 isolate of S. pseudintermedius (0.4%), whereas no isolates carried the mecC gene. 34 isolates were resistant to cefoxitin (14.6%) and they were all mecA positive. The results of this study show an MR prevalence of 64.2% confirming concerns about antibiotic resistance in veterinary medicine. In conclusion, this is the first study analyzing the prevalence of mecC and pvl in comparison to mecA, in a large cohort of CoPS clinical isolates from dogs with pyoderma. A multimodal surveillance on the prevalence of mecC and pvl in veterinary medicine is essential to appropriate antimicrobial management. [ABSTRACT FROM AUTHOR]

Background Diabetic foot ulcers (DFUs) are common complications in diabetes patients, often leading to sepsis and leg amputation. Methicillin-resistant Staphylococcus aureus (MRSA) infections in DFUs pose challenges due to methicillin resistance with mecA and mecC genes. This study aims to assess the prevalence of MRSA in clinical isolates from DFUs, analyze the antibiogram of MRSA isolates, and detect the presence of the mecA and mecC genes among MRSA isolates. Methodology The isolated S. aureus colonies were identified and antimicrobial susceptibility was performed using the Vitek-2 Compact system. Methicillin resistance was also confirmed through the disc diffusion method. Confirmed methicillin-resistant isolates were subjected to real-time polymerase chain reaction (RT-PCR) to detect mecA and mecC genes. Results A total of 474 purulent samples from DFUs yielded 541 distinct isolates, comprising 201 gram-positive and 340 gram-negative organisms. Among the gram-positive organisms, Staphylococcus species predominated, with 79 S. aureus isolates, 34 of which were methicillin-resistant. All MRSA isolates (100%) were sensitive to tetracycline, linezolid, teicoplanin, and vancomycin, and 94% were sensitive to cotrimoxazole but least susceptible to ciprofloxacin and levofloxacin. RT-PCR confirmed the presence of mecA genes in all 34 isolates and mecC genes in three isolates. Conclusions The presence of mecA in all 34 MRSA isolates underscores consistent methicillin resistance. The co-occurrence of mecA and mecC in three isolates hints at genetic diversity. Two MRSA isolates positive for mecC were isolated from rural patients involved in farming and animal husbandry, suggesting an occupational risk. The third patient was from a non-rural area, indicating potential alternative transmission pathways warranting further investigation., Competing Interests: Human subjects: Consent was obtained or waived by all participants in this study. JSS Medical College Institutional Ethics Committee issued approval JSS/MC/PG/48. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, GS et al.)

In 2011, a novel methicillin resistance gene, mecC , was described in human and bovine Staphylococcus aureus isolates. mecC- positive S. aureus is most commonly associated with livestock and wildlife populations across Europe and is particularly prevalent in hedgehogs, but only occasionally causes human infections. In this study, we characterize and investigate the origin of two human S. aureus isolates containing mecC genes from New Zealand. The two isolates were identified from patients with severe invasion infections as part of an S. aureus bacteraemia study. Whole-genome sequencing was used to characterize staphylococcal cassette chromosome mec (SCC mec ) elements and perform phylogenetic comparisons with publicly available strains from mecC -associated clonal complexes, including isolates from hedgehogs from New Zealand and Europe/United Kingdom (UK), and livestock, wildlife and human isolates from Europe/UK. The two isolates from our study have almost identical SCC mec type XI elements containing a mecC gene. However, this gene contains a premature stop codon, consistent with the methicillin-susceptible phenotype observed for these isolates. Core genome SNP analyses showed that the two isolates are 234 SNPs apart and are most closely related to an isolate obtained from a New Zealand hedgehog. However, there are considerable differences in the mecC mobile element between the human and hedgehog isolates, indicating the presence of an as-yet-unknown reservoir of mecC S. aureus in the New Zealand environment., Competing Interests: The authors declare that there are no conflicts of interest., (Copyright © 2024 The Authors.)

This study evaluated the use of the Model of Engaging Communities Collaboratively (MECC) to guide the Jandu Yani U (For All Families) project, in which the Triple P - Positive Parenting Program was collaboratively adapted for use in very remote Aboriginal communities in Western Australia. The communities' responses to the MECC processes were evaluated through interview-style or selfadministered surveys, semiformal interviews, focus group discussions and storytelling. The MECC processes were acceptable across all groups (mean score 3.86 on a 5-point acceptability scale). Qualitative data supported and gave context to the quantitative findings, demonstrating the acceptability and utility of the approach. The MECC provided a valuable framework for Aboriginal community engagement, program dissemination and implementation of research. [ABSTRACT FROM AUTHOR]

Methicillin-resistant in Staphylococci is a serious public health issue. It is mostly encoded by the mecA gene. The mecC gene is a new mecA analog responsible for resistance to methicillin in some Staphylococcal clinical isolates. This mecC gene is still underestimated in Egypt. The aim of the current study was to detect mecA and mecC genes in clinical Staphylococci isolates from a tertiary care university hospital in Egypt compared to the different phenotypic methods. A total of 118 Staphylococcus aureus (S. aureus) and 43 coagulase-negative Staphylococci (CoNS) were identified from various hospital-acquired infections. Methicillin resistance was identified genotypically using the PCR technique and phenotypically using the cefoxitin disc diffusion test, oxacillin broth microdilution and the VITEK2 system in all Staphylococcal isolates. The mecA gene was detected in 82.2% of S. aureus and 95.3% of CoNS isolates, while all of the isolates tested negative for the mecC gene. Interestingly, 30.2% of CoNS isolates showed the unique character of inducible oxacillin resistance, being mecA-positive but oxacillin-susceptible (OS-CoNS). The dual use of genotypic and phenotypic methods is highly recommended to avoid missing any genetically divergent strains.

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.

The recent emergence of LA-MRSA and their identification in people in contact with livestock has increased their importance for dairy sector and has led to the need to check MRSA presence in dairy foods. Therefore, a total of 285 dairy food samples were analyzed for the presence of MRSA. The 285 samples included raw milk (50 samples), traditional cheeses consisting of white-pickled cheese (50 samples) and Tulum cheese (50 samples), yogurt (25 samples), butter (25 samples), traditional clotted cream (20 samples), pastry cream (50 samples), and traditional Maras ice-cream (15 samples). The isolation procedure of S. aureus followed the EN/ISO 6888–1 (01/2004) method. Out of 99 presumptive isolates, 27 S. aureus isolate were identified by both MALDI-TOF MS and PCR. Overall, 15 out of 285 samples (5.26%) were contaminated by S. aureus; however, mean contamination level of S. aureus between different types of foods was observed as the highest in pastry creams with 6.4 × 103 CFU/g No S. aureus was isolated in yogurt, butter, traditional clotted cream, and traditional Maras ice-cream samples. Strains with MRSA characteristics were investigated by real-time PCR (RTiPCR)-targeting mecA and mecC genes. Out of 27 S. aureus strains, 4 strains carried one gene [mecA gene in 2 strains, isolated from raw milk (sample no: 27) and a White-pickled cheese (sample no: 46); 2 strains carried mecC gene, isolated from two different pastry creams (sample no: 33 and 44)], and only a strain isolated from a pastry cream (sample no: 21) harbored both mecA and mecC genes. None of the strains from Tulum cheeses were positive with either mec genes. This study revealed the relationship of MRSA with raw milk and traditional artisanal dairy products. Extra preventive measures to avoid MRSA-contaminated dairy foods must be applied as their consumption poses a potential risk to public health.

Nowadays, bovine mastitis has been recorded as an inflammation of the mammary gland and Staphylococcus aureus is the main pathogens responsible for contagious mastitis in ruminants. Presented study is looking for to assess the existence of methicillin-resistant and PVL positive Staphylococcus aureus from bovine milk and if there is any relation among the detected genes and antibiotic resistance or not? For this purpose, 100 milk samples were collected from cows affected with mastitis from four dairy farms in Fars province. The samples were transferred to MSA medium and incubated at 37 °C for 24 h. The pure colonies were identified by biochemical as well as molecular techniques using mecA, mecC and PVL gene. Then, the antibiotic pattern of the isolates was analyzed based on CLSI 2019 and Whonet program. Finally, the results were analyzed using statistical analysis. Totally, 6 S. aureus strains were detected which was confirmed by biochemical as well as molecular techniques. The isolates showed different antibiotic susceptibility patterns, most of the isolates were multidrug resistant, while, the strain No. 5 was the most sensitive one. Our results indicated presence of mecA gene in strains 1, 2 and 3, mecC in isolates 4, 5 and 6, and the PVL gene only in strain 4. Hence, it could be concluded that although different phages transporting PVL genes to the S. aureus isolate, in this special research we didn't find any relationship between presence of mecA, mecC, and PVL genes with antibiotics resistant pattern. [ABSTRACT FROM AUTHOR]

Although the distinction between the Community-Associated-Methicillin-Resistant Staphylococcus aureus (CA-MRSA) and Hospital-Associated-Methicillin-Resistant S. aureus (HA-MRSA) has blurred in recent years, the CA-MRSA is an important group because of its potential to cause fulminant and severe infections. Its importance has further increased with the emergence of Livestock-Associated-Methicillin-Resistant S. aureus (LA-MRSA).

Background: Staphylococcus aureus (S. aureus) is one of the leading causes of community and hospital acquired infections globally. The objective of the study was to assess the prevalence, study the carriage of antibiotic resistance genes and evaluate the molecular typing of S. aureus isolates from a tertiary care hospital in Islamabad. Methods: A total of 1528 staphylococci isolates were included in this study. Standard microbiological procedures were applied to identify S. aureus. Antimicrobial susceptability was evaluated using the disk diffusion method and Minimum Inhibitory Concentrations (MICs) were determined using microbroth dilution method following Clinical and Laboratory Standards Institute (CLSI) guidelines. Multiplex PCR was used to detect antibiotic resistance genes, and molecular typing was performed using agr, SCCmec, spa, and Multi-Locus Sequence Typing (MLST) and clonal relatedness by Pulse Field Gel Electrophoresis (PFGE) methods. Results: Overall 65% were MRSA and 35% were methicillin sensitive Staphylococcus aureus (MSSA). Among MRSA isolates, 83% were multi-drug resistant and mecA was found in 54% isolates, mecC was in 3% while 1 MRSA carried both mecA and mecC genes. agrI (22%) was most prevalent group in MRSA, while agrIII (16%) was observed in MSSA. SCCmec types I, II, III, IV, and VI were detected, with high prevalence of type III while type V was absent. The prevelant spa type in MRSA was t657 with SCCmecIII elelments while in MSSA it was t021. One NEW spa type identified in MSSA isolates. In a subset of isolates, ST772 with SCCmecIV, ST1 carrying PVL marker, and ST1535 was reported first time from Pakistan. Conclusions: The study presents a comprehensive analysis of prevalent S. aureus types and their antibiotic resistance profiles. It also reports for the first time SCCmec type VI and clinical MRSA isolates with mecC alone and in combination with mecA from Islamabad, Pakistan. This calls for further detailed investigations in other hospital settings in the region.

Methicillin-resistant Staphylococcus aureus bearing the mecC gene (mecC-MRSA) has been reported from animals and humans in recent years. This study describes the first mecC-MRSA isolates of human and equine origin in Hungary (two isolates from horses and one from a veterinarian, who treated one of the infected horses, but was asymptomatic). MRSA isolates were identified by cultivation and PCR detection of the species-specific spa gene and mecA/mecC methicillin resistance genes. The isolates were characterized by antibiotic susceptibility testing, MLST, spa, SCCmec typing, PFGE and whole genome sequencing (WGS). All three isolates belonged to the ST130-t843-SCCmec XI genotype, and carried the mecC and blaZ genes. Apart from beta-lactam drugs, they were sensitive to all tested antibiotics. The isolates of the infected horse and its veterinarian had the same PFGE pulsotype and showed only slight differences with WGS. Hence, this is the first description of direct transmission of a mecC-carrying MRSA between a horse and its veterinarian. The emergence of mecC in the country highlights the importance of the appropriate diagnostics in MRSA identification.

Mobile Edge Computing and Communication (MECC) can be deployed in close proximity with sensing devices and act as middleware between cloud and local networks. The health and fitness movement has become extremely popular recently. Endurance activities, such as marathons, triathlons, and cycling have also grown in popularity. However, with more people participating in these activities, more accidents and injuries occur—ranging from heat stroke, to heart attacks, shock, or hypoxia. All physical training activities include a risk of injury and accidents. Therefore, any research that offers a means of reducing injury risk will significantly contribute to the personal fitness field. Moreover, with the growing popularity of wearable devices and the rise of the MECC, the development and application of wearable devices that can connect to the MECC has become widespread, producing many new innovations. Although many wearable devices, such as wrist straps and smart watches, are available and able to detect individual physiological data, they cannot monitor the human body in a state of motion. Therefore, this study proposes a set of monitoring parameters for a novel wearable device connected to the MECC based on fitness management to assist fitness trainers in effective prompted strength training, and to offer timely warnings in the event of an injury risk. The data collected by the monitoring device using fuzzy theory include risk factor, body temperature, heart rate, and blood oxygen concentration. The proposed system can display in real-time the current physiological state of a wearer/user. The introduction of this device will hopefully enable trainers to immediately and effectively control and monitor the intensity of a training session, while increasing training safety, and offer crucial and immediate diagnostic information so that the correct treatment can be applied without delay in the event of injury. [ABSTRACT FROM AUTHOR]

Coagulase positive Staphylococci (CoPS) are the leading cause of canine cutaneous and otic infections. Virulence factors associated with Staphylococci include the expression of mec and panton-valentine leukocidin (pvl) genes. Methicillin-resistance (MR) is commonly associated with mecA gene expression, although a recently identified variant, mecC, has been reported. This study aims to evaluate the prevalence of mecA, mecC and pvl genes in 232 clinical isolates of CoPS collected from dogs with pyoderma. A multiplex PCR, and Kirby-Bauer disk diffusion susceptibility test for cefoxitin was performed for all isolates. PBP2a agglutination test was performed on 127 isolates. Standard MRSA isolates were used as positive controls. The mecA gene was identified in 149/232 isolates (64.2%): 116 S. pseudintermedius, 30 S. coagulans and three S. aureus. The pvl gene was present in only 1 isolate of S. pseudintermedius (0.4%), whereas no isolates carried the mecC gene. 34 isolates were resistant to cefoxitin (14.6%) and they were all mecA positive. The results of this study show an MR prevalence of 64.2% confirming concerns about antibiotic resistance in veterinary medicine. In conclusion, this is the first study analyzing the prevalence of mecC and pvl in comparison to mecA, in a large cohort of CoPS clinical isolates from dogs with pyoderma. A multimodal surveillance on the prevalence of mecC and pvl in veterinary medicine is essential to appropriate antimicrobial management.

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There is a lack of information concerning mecC clinical methicillin-resistant Staphylococcus aureus (MRSA) strains throughout the world. In the present survey, 345 MRSA strains were characterized by antimicrobial susceptibility testing and staphylococcal cassette chromosome mec element (SCCmec) typing. mecC-positive MRSA isolates were characterized by study of biofilm formability, adhesion and virulence analysis, multilocus sequence typing, accessory gene regulator (agr) typing, S. aureus protein A locus (spa) typing and staphylocoagulase typing. The present study found ten SCCmec types, with the majority being SCCmec type III (38.3%). The presence of mecC was confirmed in three isolates from skin wounds (two isolates) and burn wounds (one isolate). All the mecC-positive isolates carried SCCmec XI and belonged to coa type III. Molecular typing showed that these isolates belonged to clonal complex/ST130-spa type t843-agr type III (two isolates) and clonal complex/ST599-spa type 5930-agr type I. The presence of SCCmec type IV confirms the hypothesis of extensive infiltration from the community to the hospital. Detection of MRSA isolates harbouring the mecC gene highlights the need to perform routine detection methods and molecular investigations in order to identify these emerging strains and limit their transfer in hospitals and communities.

Background: Staphylococcus aureus(S. aureus)causes a difficult problem in the poultry industry because it causes diseases thatare difficult to treatdue to the resistance of these bacteria to antibiotics and their possession of a battery of virulence and resistance genes in addition to their ability to producethick biofilms.Method: A cross-sectional studyconducted to collecta total of 53samples from different clinical cases in broilers during the period from August 2019 to February 2020in Al-Najaf and Karbala cities,The clinical isolates were determined by using the conventional standard biochemical tests. All the specimens cultured on blood agar medium supplemented with 5% blood for primary isolation and selected by using selective mediamannitol salt agar(MSA) for confirmation the mannitol fermentation, then subjected to gram’s staining, catalase, oxidase,and further slidecoagulase test, then allS. aureusisolatestested by antibiotic susceptibility test, and screened for the presence of mecA and mecC genes using PCR for the detection of MRSA isolates, then subjected to the detection of virulence genes (pvl and eta), antibiotic resistance gene (cfr), identification of integron class 1, biofilm formation assay, themulti-druge resistance profiles(MDR)and multible antibiotics resistance (MAR)indexes werecalculated.Results: the isolation rate of S. aureusfrom the broilers'clinical samples was 37.7%. The antibiotic susceptibility test revealed that 85%of S. aureusisolateswere resistant to one or moreofthe antibiotic tested. All 53isolates were assessed for the presence of mecA and mecC genes byusingPCR. The mecA gene-specific PCRproductwas seen in 7 (35%)isolates and considered as MRSA. Among all S. aureus isolates,two isolates were positive for the eta gene, and 15 (75%) isolates harboring integron class 1, while the biofilm formation test revealed that 7 (35%) was positive biofilm producers and three of them werestrong producers,consequentlly, 13 (65%) of the isolates wereresisted to three or more antibiotics and considered as MDR strains.While pvl, cfr, and mecC gene werenot detected among S. aureus isolates.Conclusion:the current study revealed thatS. aureuspossessa real threat in the poultry industry reflectingapublic healthproblemdue to the large acquisition of antibiotic resistance genes by these bacteria,the results indicated a high percentage of isolates havingMDR characteristic, and two of them were resistant to all antibiotics tested.In addition to the presence of two MRSA isolates carrying the eta gene, this indicatingthat they are of human origin.

The recent emergence of LA-MRSA and their identification in people in contact with livestock has increased their importance for dairy sector and has led to the need to check MRSA presence in dairy foods. Therefore, a total of 285 dairy food samples were analyzed for the presence of MRSA. The 285 samples included raw milk (50 samples), traditional cheeses consisting of white-pickled cheese (50 samples) and Tulum cheese (50 samples), yogurt (25 samples), butter (25 samples), traditional clotted cream (20 samples), pastry cream (50 samples), and traditional Maras ice-cream (15 samples). The isolation procedure of S. aureus followed the EN/ISO 6888–1 (01/2004) method. Out of 99 presumptive isolates, 27 S. aureus isolate were identified by both MALDI-TOF MS and PCR. Overall, 15 out of 285 samples (5.26%) were contaminated by S. aureus; however, mean contamination level of S. aureus between different types of foods was observed as the highest in pastry creams with 6.4 × 103 CFU/g No S. aureus was isolated in yogurt, butter, traditional clotted cream, and traditional Maras ice-cream samples. Strains with MRSA characteristics were investigated by real-time PCR (RTiPCR)-targeting mecA and mecC genes. Out of 27 S. aureus strains, 4 strains carried one gene [mecA gene in 2 strains, isolated from raw milk (sample no: 27) and a White-pickled cheese (sample no: 46); 2 strains carried mecC gene, isolated from two different pastry creams (sample no: 33 and 44)], and only a strain isolated from a pastry cream (sample no: 21) harbored both mecA and mecC genes. None of the strains from Tulum cheeses were positive with either mec genes. This study revealed the relationship of MRSA with raw milk and traditional artisanal dairy products. Extra preventive measures to avoid MRSA-contaminated dairy foods must be applied as their consumption poses a potential risk to public health. [ABSTRACT FROM AUTHOR]

In this study, twelve broiler farms were randomly sampled in three different cities (Adana, Hatay and Sakarya) in Turkey, to investigate the presence and genetic diversity of methicillin resistant coagulase-negative staphylococci (MRCoNS). Four houses for each broiler flock were selected for sampling and one hundred throat swabs from each house, from each broiler flock were collected and divided into five pools of 20 swabs. Overall, 63 methicillin resistant S. sciuri were isolated, and mecA was alone detected in 56 isolates, seven isolates were positive not only for mecA, but also for mecC. Most of the isolates (77.8%) were non-typeaple for SCCmec and ten isolates (15.9%) carried SCCmec type III. All isolates were resistant to cefoxitin and penicillin, and the frequency of the isolates for trimethoprim/sulfamethoxazole, clindamycin, erythromycin, tetracycline resistance ranged between 66.7% and 73%. The PFGE analysis revealed the presence of 47 distinct pulsotypes. The most common resistant genes detected were lnuA and tet genes. None of the isolates was positive for toxin and disinfectant resistance genes. The present study is the first reporting the presence of the mecC gene among CoNS in Turkey. The results of this study also indicated that broiler flocks were potential source of multi-resistant methicillin resistant S. sciuri, with a high diversity. [ABSTRACT FROM AUTHOR]

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.

Introduction: A fast and accurate determination of methicillin resistance in Staphylococcus aureus strains is vital. This study aimed to compare the sensitivity and specificity of the cefoxitin disc diffusion (CDD) test and BD Phoenix automated system considering mecA/mecC positivity as the gold standard and to investigate the presence of Panton-Valentine leukocidin (PVL) toxin gene, a crucial virulence factor of S. aureus strains. Materials and Methods: Overall, 179 Staphylococcus aureus strains from various clinical samples were included. Antibiotic sensitivity was tested using the Phoenix automated system and by applying the Kirby-Bauer disc diffusion method for cefoxitin (30 μg). The mecA, mecC, and PVL presence was determined using the conventional multiplex polymerase chain reaction method. mecA/mecC positivity was considered as the gold standard. Statistical analysis was performed using SPSS 15.0 for Windows (Chicago, IL, USA). Results: Overall, 91 strains (50.8%) were mecA positive and identified as methicillin resistant Staphylococcus aureus (MRSA). No isolates containing the mecC gene were detected. The Phoenix automated system falsely identified six methicillin-sensitive S. aureus (MSSA) isolates, which were mecA and mecC negative as MRSA. The sensitivity and specificity of the CDD test were found to be 100% in determining MRSA, and the sensitivity and specificity the Phoenix automated system were 100% and 93.2%, respectively. The PVL positivity rate in MRSA and MSSA strains was 6.5% and 7.4%, respectively. All PVL-positive strains were isolated from the skin and soft tissues. Conclusion: The CDD test is a reliable method for routine procedures. Methicillin-sensitive strains can be determined as MRSA via the Phoenix automated system. Nevertheless, mecC-controlled MRSA should not be excluded from methods used for determining methicillin resistance. Panton-Valentine leukocidin toxin gene should be determined to enable clinicians to understand the infection severity.

Staphylococcus aureus (S. aureus) is one of the leading causes of community and hospital acquired infections globally. The objective of the study was to assess the prevalence, study the carriage of antibiotic resistance genes and evaluate the molecular typing of S. aureus isolates from a tertiary care hospital in Islamabad. A total of 1528 staphylococci isolates were included in this study. Standard microbiological procedures were applied to identify S. aureus. Antimicrobial susceptability was evaluated using the disk diffusion method and Minimum Inhibitory Concentrations (MICs) were determined using microbroth dilution method following Clinical and Laboratory Standards Institute (CLSI) guidelines. Multiplex PCR was used to detect antibiotic resistance genes, and molecular typing was performed using agr , SCC mec , spa , and Multi-Locus Sequence Typing (MLST) and clonal relatedness by Pulse Field Gel Electrophoresis (PFGE) methods. Overall 65% were MRSA and 35% were methicillin sensitive Staphylococcus aureus (MSSA). Among MRSA isolates, 83% were multi-drug resistant and mec A was found in 54% isolates, mec C was in 3% while 1 MRSA carried both mec A and mec C genes. agr I (22%) was most prevalent group in MRSA, while agr III (16%) was observed in MSSA. SCC mec types I, II, III, IV, and VI were detected, with high prevalence of type III while type V was absent. The prevelant spa type in MRSA was t657 with SCC mec III elelments while in MSSA it was t021. One NEW spa type identified in MSSA isolates. In a subset of isolates, ST772 with SCC mec IV, ST1 carrying PVL marker, and ST1535 was reported first time from Pakistan. The study presents a comprehensive analysis of prevalent S. aureus types and their antibiotic resistance profiles. It also reports for the first time SCC mec type VI and clinical MRSA isolates with mec C alone and in combination with mec A from Islamabad, Pakistan. This calls for further detailed investigations in other hospital settings in the region. [ABSTRACT FROM AUTHOR]

The aim of this study was to detect and characterize isolates of methicillin-/oxacillin-resistant Staphylococcus aureus (MRSA) carrying gene mecC (MRSA/mecC) and occurring in the Czech Republic within the period from 2002 to 2017. Altogether, 18 from 3,472 isolates of MRSA were mecC positive (0.52%). The first detection of MRSA/mecC in the Czech Republic is dated to 2004. MRSA/mecC isolates were susceptible to almost all tested antibiotics with few exceptions. Resistances to erythromycin (n = 2), clindamycin (n = 1), trimethoprim-sulfamethoxazole (n = 1), and rifampicin (n = 1) were found in the collection. Multilocus sequence typing and spa typing revealed a genetic heterogeneity of MRSA/mecC strains: three CCs (130, 425, and 2361), five STs (1245, 130, 2361, 425, and a new ST5480), and eight spa types (t843, t978, t1048, t1535, t1736, t6104, t8842, and t17153), which were detected in the study, with the highest prevalence of CC130/t843 lineage (n = 8, 44%). Except for two strains, none from 18 examined isolates harbored genes encoding any of S. aureus toxins: enterotoxins a–u, exfoliative toxins A, B, and D, toxic shock syndrome toxin-1, and the Panton-Valentine leukocidin. [ABSTRACT FROM AUTHOR]

Introduction: Methicillin-resistant Staphylococcus aureus (MRSA) is associated with adverse clinical outcomes and increased morbidity, mortality, length of hospital stay, and health-care costs. Rapid diagnosis of MRSA infections has been associated with positive impact on clinical outcomes.Areas Covered: We searched relevant papers in PubMed for the last 10 years. In major papers, we scanned the bibliographies to ensure that important articles were included. This review describes screening and diagnostic test methods for MRSA and their analytical performances with a focus on rapid molecular-based assays including those that are on the horizon. Future novel technologies will allow more rapid detection of phenotypic resistance. In the case of whole-genome sequencing, detection of mutations may predict resistance, transmission, and virulence.Expert Opinion: Currently there are many diagnostic options for the detection of MRSA in surveillance and clinical samples. In general, these are highly accurate and have resulted in improvements in targeted management and reduction in hospital or intensive care unit length of stay for both MSSA and MRSA. Impact on mortality has been variable. Promising novel technologies will not only accurately identify pathogens and detect their resistance markers but will allow discovery of virulence determinants that might further affect patient management. [ABSTRACT FROM AUTHOR]

Background: Staphylococcus aureus (S. aureus) causes a difficult problem in the poultry industry because it causes diseases that are difficult to treat due to the resistance of these bacteria to antibiotics and their possession of a battery of virulence and resistance genes in addition to their ability to produce thick biofilms. Method: A cross-sectional study conducted to collect a total of 53 samples from different clinical cases in broilers during the period from August 2019 to February 2020 in Al-Najaf and Karbala cities, The clinical isolates were determined by using the conventional standard biochemical tests. All the specimens cultured on blood agar medium supplemented with 5% blood for primary isolation and selected by using selective media mannitol salt agar (MSA) for confirmation the mannitol fermentation, then subjected to gram's staining, catalase, oxidase, and further slide coagulase test, then all S. aureus isolates tested by antibiotic susceptibility test, and screened for the presence of mecA and mecC genes using PCR for the detection of MRSA isolates, then subjected to the detection of virulence genes (pvl and eta), antibiotic resistance gene (cfr), identification of integron class 1, biofilm formation assay, the multidruge resistance profiles (MDR) and multible antibiotics resistance (MAR) indexes were calculated. Results: the isolation rate of S. aureus from the broilers' clinical samples was 37.7%. The antibiotic susceptibility test revealed that 85% of S. aureus isolates were resistant to one or more of the antibiotic tested. All 53 isolates were assessed for the presence of mecA and mecC genes by using PCR. The mecA gene-specific PCR product was seen in 7 (35%) isolates and considered as MRSA. Among all S. aureus isolates, two isolates were positive for the eta gene, and 15 (75%) isolates harboring integron class 1, while the biofilm formation test revealed that 7 (35%) was positive biofilm producers and three of them were strong producers, consequentlly, 13 (65%) of the isolates were resisted to three or more antibiotics and considered as MDR strains. While pvl, cfr, and mecC gene were not detected among S. aureus isolates. Conclusion: the current study revealed that S. aureus possess a real threat in the poultry industry reflecting a public health problem due to the large acquisition of antibiotic resistance genes by these bacteria, the results indicated a high percentage of isolates having MDR characteristic, and two of them were resistant to all antibiotics tested. In addition to the presence of two MRSA isolates carrying the eta gene, this indicating that they are of human origin. [ABSTRACT FROM AUTHOR]

Aims: Livestock‐associated methicillin‐resistant Staphylococcus aureus (LA‐MRSA) is an important public health problem in many countries. Despite reports of such isolates being found in both animals and humans in the United Kingdom few data are available on the prevalence in humans of such isolates. A prevalence study was therefore undertaken in the north‐west of England. Methods and Results: One thousand two hundred and forty‐two human MRSA isolates collected during 2015 were screened by PCR to detect two of the major forms of LA‐MRSA: clonal complex (CC)398 and mecC MRSA. Isolates identified were further characterized by antimicrobial susceptibility testing and whole‐genome sequencing using HiSeq technology. A single mecC MRSA isolate and three CC398 LA‐MRSA were identified among the isolates screened. All four isolates were from MRSA screening. A phylogenetic analysis, including previously sequenced isolates from the United Kingdom, provided strong evidence for the genomic and epidemiological linkage between a pair of animal and human isolates of CC398 LA‐MRSA in England. These data are indicative of animal and humans isolates of CC398 LA‐MRSA being involved in the same transmission network and is the first demonstration of such closely linked animal and human isolates of this lineage in the UK. Conclusions: The study indicates there is a low prevalence of CC398 LA‐MRSA and mecC MRSA among MRSA isolates in the sampled population. Significance and Impact of the Study: While the study demonstrates that LA‐MRSA were rare among human MRSA isolates in the sampled population, the data provide a baseline for the future surveillance of what is a significant public health challenge in some regions. The demonstration of linked human and animal isolates of CC398 LA‐MRSA, supported by genomic and epidemiological data, reinforces the need for such surveillance and for continued awareness of the risks that LA‐MRSA may pose in the UK. [ABSTRACT FROM AUTHOR]

Natural aquatic environments represent one of the most important vehicles of bacterial dissemination. Therefore, we aimed to isolate staphylococci from surface waters and to investigate the presence of antimicrobial resistance genes and virulence factors as well as the genetic lineages of all Staphylococcus aureus isolates. Staphylococci were recovered from water samples collected from 78 surface waters, including rivers, streams, irrigation ditches, dams, lakes, and fountains. The presence of antimicrobial resistance genes and virulence factors was investigated by PCR. Multilocus sequence typing and spa-typing were performed in all S. aureus isolates. From the 78 water samples, 33 S. aureus, one S. pseudintermedius, and 51 coagulase-negative staphylococci (CoNS) were identified. Among the S. aureus isolates, four MRSA were identified, and all harbored the mecC gene. Fourteen S. aureus were susceptible to all antimicrobials tested and the remaining showed resistance to penicillin, erythromycin and/or tetracycline encoded by the blaZ, ermT, msr(A/B), tetL, and vgaA genes. Regarding the clonal lineages, one mecC-MRSA isolate belonged to spa-type t843 and sequence type (ST) 130 and the other three to t742 and ST425. The remaining S. aureus were ascribed 14 spa-types and 17 sequence types. Eleven species of CoNS were isolated: S. sciuri, S. lentus, S. xylosus, S. epidermidis, S. cohnii spp. urealyticus, S. vitulinus, S. caprae, S. carnosus spp. Carnosus, S. equorum, S. simulans, and S. succinus. Thirteen CoNS isolates had a multidrug resistance profile and carried the following genes: mecA, msr(A/B), mph(C), aph(3′)-IIIa, aac(6′)-Ie–aph(2′’)-Ia, dfrA, fusB, catpC221, and tetK. A high diversity of staphylococci was isolated from surface waters including mecCMRSA strains and isolates presenting multidrug-resistance profiles. Studies on the prevalence of antibiotic-resistant staphylococci in surface waters are still very scarce but extremely important to estimate the contribution of the aquatic environment in the spread of these bacteria.

Mobile Edge Computing and Communication (MECC) can be deployed in close proximity with sensing devices and act as middleware between cloud and local networks. The health and fitness movement has become extremely popular recently. Endurance activities, such as marathons, triathlons, and cycling have also grown in popularity. However, with more people participating in these activities, more accidents and injuries occur—ranging from heat stroke, to heart attacks, shock, or hypoxia. All physical training activities include a risk of injury and accidents. Therefore, any research that offers a means of reducing injury risk will significantly contribute to the personal fitness field. Moreover, with the growing popularity of wearable devices and the rise of the MECC, the development and application of wearable devices that can connect to the MECC has become widespread, producing many new innovations. Although many wearable devices, such as wrist straps and smart watches, are available and able to detect individual physiological data, they cannot monitor the human body in a state of motion. Therefore, this study proposes a set of monitoring parameters for a novel wearable device connected to the MECC based on fitness management to assist fitness trainers in effective prompted strength training, and to offer timely warnings in the event of an injury risk. The data collected by the monitoring device using fuzzy theory include risk factor, body temperature, heart rate, and blood oxygen concentration. The proposed system can display in real-time the current physiological state of a wearer/user. The introduction of this device will hopefully enable trainers to immediately and effectively control and monitor the intensity of a training session, while increasing training safety, and offer crucial and immediate diagnostic information so that the correct treatment can be applied without delay in the event of injury.

The possible spillover from pigs into other production animals incites concern for unresolved reservoirs of human exposure. The present investigation was therefore initiated, to elucidate if Danish veal and dairy farms constitute a reservoir of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) CC398 and to potentially identify the source of introduction. We collected nasal swab samples from 17 Danish veal farms, 2 slaughterhouses, and received bulk tank milk samples from 286 dairy farms. All samples were analyzed by culturing and screening on MRSA selective plates and presumed MRSA was verified by MALDI-TOF and PCR. MRSA isolates were subjected to spa typing and whole-genome sequencing. LA-MRSA was found on two veal farms in one and three calves, respectively, with subsequent follow-up samples found negative. Eight of 286 dairy farms (2.8%) were found LA-MRSA positive and follow-up samples, from five farms showed intermittent detection of LA-MRSA. The spa types, t034 and t011, were the most common while a single isolate from a dairy farm belonged to spa type t843 associated to mecC-MRSA CC130 and is the first report of mecC-MRSA in the Danish dairy production. A phylogenetic analysis showed that some of the isolates grouped within or close to the dominant Danish pig clusters, suggesting spillover into cattle farms. Other isolates clustered outside the dominant pig clusters suggesting that other routes of introduction cannot be excluded. Results of the investigation indicated a contamination of veal farms while some dairy farms seemed to be a permanent reservoir. Thus, Danish cattle represent a low prevalence reservoir of LA-MRSA CC398, which at present, is not of major human health concern.

Background: Methicillin resistance in Staphylococcus aureus (MRSA) is classically conferred by the acquisition of the mecA gene encoding an additional penicillin binding protein with low affinity for beta-lactams. A mecA variant, named mecC, was described in 2011. MRSA isolates harboring mecC of both animal and human origin have since been collected in different European countries. In France, animal cases were reported in 4 dairy farms between 2008 and 2013 in the Meurthe-et-Moselle county, all located in a 30 km perimeter, suggesting a possible dissemination of mecC-positive MRSA strains. We performed a prospective study to evaluate the local epidemiology of such strains in terms of (i) dissemination among animals, humans and in the environment, and (ii) persistence in Meurthe-et-Moselle dairy cattle farms.Methods: The 4 French dairy farms with previous reports of mecC-positive MRSA strains and 14 farms in the same perimeter were included in this study. In each farm, nasal swabs, rectal swabs and milk samples were collected from 10 randomly selected cows, as well as nasal samples from family pets, volunteer farmers and veterinarians. One farm (E0), in which mecC-MRSA isolates were detected, was selected to study more deeply the dissemination of mecC-positive strains within the farm. After pre-enrichment of swabs and milk, they were subcultured on MSSA/MRSA chromogenic selective agar plates. S. aureus colonies were tested with a multiplex PCR to detect the mecA and mecC genes. The mecC-positive strains were characterized using DNA microarray.Results:mecC-positive strains were recovered in four farms, corresponding to the ones with previous reports of mecC-positive MRSA strains, and originated only from dairy cow samples. The screening in the E0 farm showed that 22% of the dairy cows carried mecC-positive MRSA. Three strains were also isolated from the environmental samples. All mecC-positive strains belonged to the clonal complex CC130 and harbored the same spa-type t1736.Conclusion: This study found that mecC-positive MRSA isolates are able to persist within the same farms for several years after being introduced in this setting and are able to widely disseminate but only among dairy cows suggesting that milking machines might be a key player.

The frequent carriage of Staphylococcus aureus, including methicillin-resistant S. aureus (MRSA), by wild animals along with its zoonotic potential poses a public health problem. Furthermore, the repeated detection of the mecA gene homologue, mecC, in wildlife raises the question whether these animals may be a reservoir for mecC-MRSA. Thus, we aimed to isolate S. aureus and MRSA from wild rodents living in port areas and to characterize their antimicrobial resistance and genetic lineages. Mouth and rectal swab samples were recovered from 204 wild rodents. The samples were incubated in BHI broth with 6.5% of NaCl and after 24 h at 37 °C the inoculum was seeded onto Baird-Parker agar, Mannitol Salt agar and ORSAB (supplemented with 2 mg/L of oxacillin) plates. Species identification was confirmed by MALDI-TOF MS. The antimicrobial susceptibility testing was performed by the Kirby–Bauer disc diffusion method against 14 antibiotics. The presence of virulence and resistance genes was performed by PCR. The immune evasion cluster (IEC) system was investigated in all S. aureus. All isolates were characterized by MLST, spa- and agr typing. From 204 samples, 38 S. aureus were isolated of which six MRSA were detected. Among the six MRSA isolates, three harbored the mecC gene and the other three, the mecA gene. All mecC-MRSA isolates were ascribed to sequence type (ST) 1945 (which belongs to CC130) and spa-type t1535 whereas the mecA isolates belonged to ST22 and ST36 and spa-types t747 and t018. Twenty-five S. aureus were susceptible to all antibiotics tested. S. aureus isolates were ascribed to 11 MLST and 12 spa-types. S. aureus presents a great diversity of genetic lineages in wild rodents. This is the first report of mecC-MRSA in Portugal.

The possible spillover from pigs into other production animals incites concern for unresolved reservoirs of human exposure. The present investigation was therefore initiated, to elucidate if Danish veal and dairy farms constitute a reservoir of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) CC398 and to potentially identify the source of introduction. We collected nasal swab samples from 17 Danish veal farms, 2 slaughterhouses, and received bulk tank milk samples from 286 dairy farms. All samples were analyzed by culturing and screening on MRSA selective plates and presumed MRSA was verified by MALDI-TOF and PCR. MRSA isolates were subjected to spa typing and whole-genome sequencing. LA-MRSA was found on two veal farms in one and three calves, respectively, with subsequent follow-up samples found negative. Eight of 286 dairy farms (2.8%) were found LA-MRSA positive and follow-up samples, from five farms showed intermittent detection of LA-MRSA. The spa types, t034 and t011, were the most common while a single isolate from a dairy farm belonged to spa type t843 associated to mecC -MRSA CC130 and is the first report of mecC -MRSA in the Danish dairy production. A phylogenetic analysis showed that some of the isolates grouped within or close to the dominant Danish pig clusters, suggesting spillover into cattle farms. Other isolates clustered outside the dominant pig clusters suggesting that other routes of introduction cannot be excluded. Results of the investigation indicated a contamination of veal farms while some dairy farms seemed to be a permanent reservoir. Thus, Danish cattle represent a low prevalence reservoir of LA-MRSA CC398, which at present, is not of major human health concern. [ABSTRACT FROM AUTHOR]

• 90 MRSA isolates from companion animals were examined using DNA microarray based assays and whole-genome sequencing. • Eight sequence types (STs - ST398, ST5275 (new ST), ST225, ST8, ST22, ST152, ST1, and ST45) were detected. • Although ST398 isolates remained predominant, increasing isolation rates of human-associated MRSA clones were observed in cats and dogs. The aim of the present study was to investigate the diversity of methicillin-resistant Staphylococcus aureus (MRSA) that originated from Austrian companion animals during the last five-year period. A total of 90 non-repetitive MRSA isolates were obtained during diagnostic activities from autumn 2013 to autumn 2018. They originated from horses (n = 62), cats (n = 13), dogs (n = 10), rabbits (n = 2), a domestic canary, a zoo-kept hammer-headed bat (Hypsignathus monstrosus) and a semi-captive northern bald ibis (Geronticus eremita). Antimicrobial susceptibility testing was performed. All isolates were mecA -positive and mecC -negative. The isolates were genotyped by SCC mec, spa and dru typing, Multiple-Locus Variable number of tandem repeat Analyses (MLVA), S. aureus DNA microarray, and whole-genome sequencing (WGS). Eight sequence types (STs - ST398, ST5275 (new ST), ST225, ST8, ST22, ST152, ST1, and ST45), three SCC mec types (II, IV, and V), sixteen spa types (t003, t008, t011, t015, t032, t034, t1381, t1928, t1985, t223, t334, t355, t430, t6447, t6867, and t7105), fourteen dru types (dt10a, dt10az, dt10q, dt10r, dt11a, dt5e, dt6j, dt9a, dt9ak, dt9g, and four new types dt8as, dt7ak, dt4j, dt14n), and thirty-five MLVA types were detected. WGS-based core genome MLST (cgMLST) displayed five main clusters. Compared to the time period 2004–2013, the results of the present study show not only a higher diversity among the MRSA isolates within the population of Austrian companion animals, but also the introduction of new clones. Although ST398 isolates remained predominant, mainly due to high presence of this lineage among horses, increasing isolation rates of human-associated MRSA clones were observed in cats and dogs. [ABSTRACT FROM AUTHOR]

mecA is a predefined gene causing methicillin resistance in Staphylococcus aureus (S. aureus) isolates; however, it has been shown that some methicillin-resistant S. aureus (MRSA) strains do not carry this gene. Recently, in isolates found to be MRSA-positive but mecA -negative, a new resistance gene called mecC , which is a homolog of mecA , has been reported. This study aimed to investigate the mecC and mecA genes in MRSA strains isolated from different geographic regions in Turkey. The sample of the study consisted of 494 MRSA strains isolated from seven geographical regions in Turkey between 2013 and 2016. The strains were obtained from 17 centers, comprising 13 university hospitals, three education and research hospitals, and one state hospital. Methicillin resistance in S. aureus strains was determined using the agar disk diffusion method with a cefoxitin disk and the agar dilution method with oxacillin. The mecC and mecA genes in MRSA strains was investigated by Polymerase Chain Reaction (PCR). Of the MRSA strains investigated, 47.9% were isolated from intensive care units. Concerning sample type, 36.7% were detected in the respiratory tract (tracheal aspirate, sputum, etc.), 24.8% in blood, 18.7% in skin and soft tissues, 9.3% in nasal swabs, 5.4% in urine, 4.1% in ears, and 1% in sterile body fluid. Using PCR, mecC was not identified in any of the S. aureus strains isolated from different clinical microbiology laboratories. mecA gene positivity was found in 315 of the MRSA strains (63.8%). Staphylococcal Cassette Chromosome mec (SCC mec) type was identified in 232 strains (46.9%), of which 136 (58.7%) were type II, 75 (32.4%) were type IV, 12 (5.1%) were type IIIb, six (2.5%) were type I, and three (1.3%) were type III. This is the first multi-centered study to investigate MRSA strains isolated from different regions in Turkey. The mecC gene was not detected in any of the MRSA strains. We believe that this study will constitute an important basis for monitoring possible future changes. [ABSTRACT FROM AUTHOR]

Highlights • Description of mecC -carrying CoNS from 4299 wild and domesticated animals in Austria. • First detection of mecC -positive S. caprae and S. warneri from animals. • First observation of mecC -positive CoNS isolates from companion animals. Abstract The presence of the methicillin resistance gene mecC in coagulase-negative Staphylococcus spp. (CoNS) is scarce. The aim of this study was to characterize mecC -positive CoNS isolated from various wild and domestic animals. The presence of the mecC gene was screened in 4299 samples from wild animals and domestic animals. Fifteen coagulase-negative staphylococci, that displayed a cefoxitin-resistant phenotype, were tested mecC -positive by PCR. Antimicrobial susceptibility testing was performed for all isolates. The 15 isolates were genotyped by sequencing of the entire class E mec gene complex (blaZ-mecC-mecR1-mecI), the ccrA and ccrB recombinase genes and other determinants within the type XI SCC mec element. DNA microarray analysis was performed and five selected isolates were additionally whole genome sequenced and analyzed. S. stepanovicii (n = 3), S. caprae (n = 1), S. warneri (n = 1), S. xylosus (n = 1) and S. sciuri (n = 9) were detected. All but the S. sciuri isolates were found to be susceptible to all non-beta lactams. The entire class E mec gene complex was detected in all isolates but ccrA and ccrB genes were not identified in S. stepanovicii and S. xylosus. The genes erm (B) and fexA (n = 4, each) were the most predominant non-beta lactam resistance genes detected in the S. sciuri isolates. Even though the presence of the mecC gene among CoNS is a rare observation, this study further expands our knowledge by showing that the mecC gene, including its allotypes, are present in more staphylococcal species from different animal species than has been previously described. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus is an opportunist pathogen that is responsible for numerous types of infections. S. aureus is known for its ability to easily acquire antibiotic resistance determinants. Methicillin-resistant S. aureus (MRSA) is a leading cause of infections both in humans and animals and is usually associated with a multidrug-resistant profile. MRSA dissemination is increasing due to its capability of establishing new reservoirs and has been found in humans, animals and the environment. Despite the fact that the information on the incidence of MRSA in the environment and, in particular, in wild animals, is scarce, some studies have reported the presence of these strains among wildlife with no direct contact with antibiotics. This shows a possible transmission between species and, consequently, a public health concern. The aim of this review is to better understand the distribution, prevalence and molecular lineages of MRSA in European free-living animals.

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.

ABSTRACT Typing 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.