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1

Lekkerkerk, W. S. N., W. J. B. van Wamel, S. V. Snijders, R. J. Willems, E. van Duijkeren, E. M. Broens, J. A. Wagenaar, J. A. Lindsay, and M. C. Vos. "What Is the Origin of Livestock-Associated Methicillin-Resistant Staphylococcus aureus Clonal Complex 398 Isolates from Humans without Livestock Contact? An Epidemiological and Genetic Analysis." Journal of Clinical Microbiology 53, no. 6 (March 25, 2015): 1836–41. http://dx.doi.org/10.1128/jcm.02702-14.

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Fifteen percent of all methicillin-resistantStaphylococcus aureus(MRSA) clonal complex 398 (CC398) human carriers detected in The Netherlands had not been in direct contact with pigs or veal calves. To ensure low MRSA prevalence, it is important to investigate the likely origin of this MRSA of unknown origin (MUO). Recently, it was shown that CC398 strains originating from humans and animals differ in the presence of specific mobile genetic elements (MGEs). We hypothesized that determining these specific MGEs in MUO isolates and comparing them with a set of CC398 isolates of various known origin might provide clues to their origin. MUO CC398 isolates were compared to MRSA CC398 isolates obtained from humans with known risk factors, a MRSA CC398 outbreak isolate, livestock associated (LA) MRSA CC398 isolates from pigs, horses, chickens, and veal calves, and five methicillin-susceptibleStaphylococcus aureus(MSSA) CC398 isolates of known human origin. All strains werespatyped, and the presence or absence of,scn,chp, φ3int, φ6int, φ7int,rep7,rep27, andcadDXwas determined by PCRs. The MRSA CC398 in humans, MUO, or MRSA of known origin (MKO) resembled MRSA CC398 as found in pigs and not MSSA CC398 as found in humans. The distinct human MSSA CC398spatype, t571, was not present among our MRSA CC398 strains; MRSA CC398 was tetracycline resistant and carried no φ3 bacteriophage withscnandchp. We showed by simple PCR means that human MUO CC398 carriers carried MRSA from livestock origin, suggestive of indirect transmission. Although the exact transmission route remains unknown, direct human-to-human transmission remains a possibility as well.
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2

Bouiller, Kevin, Xavier Bertrand, Didier Hocquet, and Catherine Chirouze. "Human Infection of Methicillin-Susceptible Staphylococcus aureus CC398: A Review." Microorganisms 8, no. 11 (November 5, 2020): 1737. http://dx.doi.org/10.3390/microorganisms8111737.

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Staphylococcus aureus (SA) belonging to the clonal complex 398 (CC398) took a special place within the species due to its spread throughout the world. SA CC398 is broadly separated in two subpopulations: livestock-associated methicillin-resistant SA (MRSA) and human-associated methicillin-susceptible SA (MSSA). Here, we reviewed the global epidemiology of SA CC398 in human clinical infections and focused on MSSA CC398. The last common ancestor of SA CC398 was probably a human-adapted prophage φSa3-positive MSSA CC398 strain, but the multiple transmissions between human and animal made its evolution complex. MSSA and MRSA CC398 had different geographical evolutions. Although MSSA was present in several countries all over the world, it was mainly reported in China and in France with a prevalence about 20%. MSSA CC398 was frequently implicated in severe infections such as bloodstream infections, endocarditis, and bone joint infections whereas MRSA CC398 was mainly reported in skin and soft tissue. The spread of the MSSA CC398 clone is worldwide but with a heterogeneous prevalence. The prophage φSa3 played a crucial role in the adaptation to the human niche and in the virulence of MSSA CC398. However, the biological features that allowed the recent spread of this lineage are still far from being fully understood.
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3

Ceballos, Sara, Carmen Aspiroz, Laura Ruiz-Ripa, Esteban Reynaga, José Manuel Azcona-Gutiérrez, Antonio Rezusta, Cristina Seral, et al. "Epidemiology of MRSA CC398 in hospitals located in Spanish regions with different pig-farming densities: a multicentre study." Journal of Antimicrobial Chemotherapy 74, no. 8 (May 16, 2019): 2157–61. http://dx.doi.org/10.1093/jac/dkz180.

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Abstract Background Tetracycline resistance (TetR) is a marker of livestock-associated MRSA of lineage CC398. Objectives To determine the MRSA CC398 prevalence among TetR-MRSA recovered in Spanish hospitals located in regions with different pig-farming densities, and the influence of pig density as a key risk factor for its acquisition. Methods TetR-MRSA isolates (n = 232) recovered from clinical and epidemiological samples during January–June 2016 in 20 hospitals in 13 regions with different pig-farming densities were analysed. MRSA CC398 identification, detection of spa types, methicillin resistance genes and immune evasion cluster (IEC) genes were performed by PCR/sequencing. Statistical analyses were performed to establish the relationships between MRSA CC398 prevalence and pig density. Results The global MRSA prevalence was 29.7% (6.9% TetR-MRSA/MRSA), with 137 CC398 isolates recovered, representing 4.1% of total MRSA and 59.1% of TetR-MRSA. Among MRSA CC398, 16 different spa types were recorded (t011: 72.3%), and all but two strains were IEC negative. Higher pig-density regions were associated with significant MRSA CC398 increases in hospitals located in adjacent regions (P < 0.001). Linear regression models explained the relationships between MRSA CC398 and pig density (P < 0.001), with an increase of 6.6 MRSA CC398 cases per 100 MRSA per increase of 100 pigs/km2 in a region. Conclusions High pig density leads to a significant increase in MRSA CC398 in hospitals in Spain, and its combination with a high human population could help its dissemination. In Spain, the prevalence of the zoonotic CC398 lineage is closely related to pig-farming density; therefore, specific tools could be implemented in order to detect its dissemination.
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4

Landt, Stephen G., Joseph A. Lesley, Leticia Britos, and Lucy Shapiro. "CrfA, a Small Noncoding RNA Regulator of Adaptation to Carbon Starvation in Caulobacter crescentus." Journal of Bacteriology 192, no. 18 (July 2, 2010): 4763–75. http://dx.doi.org/10.1128/jb.00343-10.

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ABSTRACT Small noncoding regulatory RNAs (sRNAs) play a key role in the posttranscriptional regulation of many bacterial genes. The genome of Caulobacter crescentus encodes at least 31 sRNAs, and 27 of these sRNAs are of unknown function. An overexpression screen for sRNA-induced growth inhibition along with sequence conservation in a related Caulobacter species led to the identification of a novel sRNA, CrfA, that is specifically induced upon carbon starvation. Twenty-seven genes were found to be strongly activated by CrfA accumulation. One-third of these target genes encode putative TonB-dependent receptors, suggesting CrfA plays a role in the surface modification of C. crescentus, facilitating the uptake of nutrients during periods of carbon starvation. The mechanism of CrfA-mediated gene activation was investigated for one of the genes predicted to encode a TonB-dependent receptor, CC3461. CrfA functions to stabilize the CC3461 transcript. Complementarity between a region of CrfA and the terminal region of the CC3461 5′-untranslated region (5′-UTR) and also the behavior of a deletion of this region and a site-specific base substitution and a 3-base deletion in the CrfA complementary sequence suggest that CrfA binds to a stem-loop structure upstream of the CC3461 Shine-Dalgarno sequence and stabilizes the transcript.
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5

Ward, M. J., C. L. Gibbons, P. R. McAdam, B. A. D. van Bunnik, E. K. Girvan, G. F. Edwards, J. R. Fitzgerald, and M. E. J. Woolhouse. "Time-Scaled Evolutionary Analysis of the Transmission and Antibiotic Resistance Dynamics of Staphylococcus aureus Clonal Complex 398." Applied and Environmental Microbiology 80, no. 23 (September 19, 2014): 7275–82. http://dx.doi.org/10.1128/aem.01777-14.

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ABSTRACTStaphylococcus aureusclonal complex 398 (CC398) is associated with disease in humans and livestock, and its origins and transmission have generated considerable interest. We performed a time-scaled phylogenetic analysis of CC398, including sequenced isolates from the United Kingdom (Scotland), along with publicly available genomes. Using state-of-the-art methods for mapping traits onto phylogenies, we quantified transitions between host species to identify sink and source populations for CC398 and employed a novel approach to investigate the gain and loss of antibiotic resistance in CC398 over time. We identified distinct human- and livestock-associated CC398 clades and observed multiple transmissions of CC398 from livestock to humans and between countries, lending quantitative support to previous reports. Of note, we identified a subclade within the livestock-associated clade comprised of isolates from hospital environments and newborn babies, suggesting that livestock-associated CC398 is capable of onward transmission in hospitals. In addition, our analysis revealed significant differences in the dynamics of resistance to methicillin and tetracycline related to contrasting historical patterns of antibiotic usage between the livestock industry and human medicine. We also identified significant differences in patterns of gain and loss of different tetracycline resistance determinants, which we ascribe to epistatic interactions between the resistance genes and/or differences in the modes of inheritance of the resistance determinants.
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6

Domínguez, Andrea Vila, Marta Carretero Ledesma, Carmen Infante Domínguez, José Miguel Cisneros, Jose A. Lepe, and Younes Smani. "In Vitro and In Vivo Virulence Study of Listeria monocytogenes Isolated from the Andalusian Outbreak in 2019." Tropical Medicine and Infectious Disease 8, no. 1 (January 12, 2023): 58. http://dx.doi.org/10.3390/tropicalmed8010058.

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In 2019, the biggest listeriosis outbreak by Listeria monocytogenes (Lm) in the South of Spain was reported, resulting in the death of three patients from 207 confirmed cases. One strain, belonging to clonal complex 388 (Lm CC388), has been isolated. We aimed to determine the Lm CC388 virulence in comparison with other highly virulent clones such as Lm CC1 and Lm CC4, in vitro and in vivo. Four L. monocytogenes strains (Lm CC388, Lm CC1, Lm CC4 and ATCC 19115) were used. Attachment to human lung epithelial cells (A549 cells) by these strains was characterized by adherence and invasion assays. Their cytotoxicities to A549 cells were evaluated by determining the cells viability. Their hemolysis activity was determined also. A murine intravenous infection model using these was performed to determine the concentration of bacteria in tissues and blood. Lm CC388 interaction with A549 cells is non-significantly higher than that of ATCC 19115 and Lm CC1, and lower than that of Lm CC4. Lm CC388 cytotoxicity is higher than that of ATCC 19115 and Lm CC1, and lower than that of Lm CC4. Moreover, Lm CC388 hemolysis activity is lower than that of the Lm CC4 strain, and higher than that of Lm CC1. Finally, in the murine intravenous infection model by Lm CC388, higher bacterial loads in tissues and at similar levels of Lm CC4 were observed. Although a lower rate of mortality of patients during the listeriosis outbreak in Spain in 2019 has been reported, the Lm CC388 strain has shown a greater or similar pathogenicity level in vitro and in an animal model, like Lm CC1 and Lm CC4.
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7

Larsen, Jesper, Julie Clasen, Julie E. Hansen, Wilhelm Paulander, Andreas Petersen, Anders R. Larsen, and Dorte Frees. "Copresence oftet(K) andtet(M) in Livestock-Associated Methicillin-Resistant Staphylococcus aureus Clonal Complex 398 Is Associated with Increased Fitness during Exposure to Sublethal Concentrations of Tetracycline." Antimicrobial Agents and Chemotherapy 60, no. 7 (May 9, 2016): 4401–3. http://dx.doi.org/10.1128/aac.00426-16.

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ABSTRACTThe tetracycline resistance genetet(K) was shown to be integrated within the predominant staphylococcal cassette chromosomemec(SCCmec) element of Danish livestock-associated methicillin-resistantStaphylococcus aureusCC398 (LA-MRSA CC398). These LA-MRSA CC398 isolates already possessedtet(M), but the acquisition oftet(K) significantly improved their fitness at sublethal concentrations of tetracycline. Becausetet(K) is genetically linked to SCCmec, the use of tetracycline in food animals may have contributed to the successful spread of LA-MRSA CC398.
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8

Laumay, Floriane, Hugo Benchetrit, Anna-Rita Corvaglia, Nathalie van der Mee-Marquet, and Patrice François. "The Staphylococcus aureus CC398 Lineage: An Evolution Driven by the Acquisition of Prophages and Other Mobile Genetic Elements." Genes 12, no. 11 (October 30, 2021): 1752. http://dx.doi.org/10.3390/genes12111752.

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Among clinically relevant lineages of Staphylococcus aureus, the lineage or clonal complex 398 (CC398) is of particular interest. Strains from this lineage were only described as livestock colonizers until 2007. Progressively, cases of infection were reported in humans in contact with farm animals, and now, CC398 isolates are increasingly identified as the cause of severe infections even in patients without any contact with animals. These observations suggest that CC398 isolates have spread not only in the community but also in the hospital setting. In addition, several recent studies have reported that CC398 strains are evolving towards increased virulence and antibiotic resistance. Identification of the origin and emergence of this clonal complex could probably benefit future large-scale studies that aim to detect sources of contamination and infection. Current evidence indicates that the evolution of CC398 strains towards these phenotypes has been driven by the acquisition of prophages and other mobile genetic elements. In this short review, we summarize the main knowledge of this major lineage of S. aureus that has become predominant in the human clinic worldwide within a single decade.
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9

Abdullahi, Idris Nasir, Carmen Lozano, Laura Ruiz-Ripa, Rosa Fernández-Fernández, Myriam Zarazaga, and Carmen Torres. "Ecology and Genetic Lineages of Nasal Staphylococcus aureus and MRSA Carriage in Healthy Persons with or without Animal-Related Occupational Risks of Colonization: A Review of Global Reports." Pathogens 10, no. 8 (August 8, 2021): 1000. http://dx.doi.org/10.3390/pathogens10081000.

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In this conceptual review, we thoroughly searched for appropriate English articles on nasal staphylococci carriage among healthy people with no reported risk of colonization (Group A), food handlers (Group B), veterinarians (Group C), and livestock farmers (Group D) published between 2000 and 2021. Random-effects analyses of proportions were performed to determine the pooled prevalence of S. aureus, MRSA, MRSA-CC398, and MSSA-CC398, as well as the prevalence of PVL-positive S. aureus from all eligible studies. A total of 166 eligible papers were evaluated for Groups A/B/C/D (n = 58/31/26/51). The pooled prevalence of S. aureus and MRSA in healthy humans of Groups A to D were 15.9, 7.8, 34.9, and 27.1%, and 0.8, 0.9, 8.6, and 13.5%, respectively. The pooled prevalence of MRSA-CC398 nasal carriage among healthy humans was as follows: Group A/B (<0.05%), Group C (1.4%), Group D (5.4%); and the following among Group D: pig farmers (8.4%) and dairy farmers (4.7%). The pooled prevalence of CC398 lineage among the MSSA and MRSA isolates from studies of the four groups were Group A (2.9 and 6.9%), B (1.5 and 0.0%), C (47.6% in MRSA), and D (11.5 and 58.8%). Moreover, MSSA-CC398 isolates of Groups A and B were mostly of spa-t571 (animal-independent clade), while those of Groups C and D were spa-t011 and t034. The MRSA-CC398 was predominately of t011 and t034 in all the groups (with few other spa-types, livestock-associated clades). The pooled prevalence of MSSA and MRSA isolates carrying the PVL encoding genes were 11.5 and 9.6% (ranges: 0.0–76.9 and 0.0–28.6%), respectively. Moreover, one PVL-positive MSSA-t011-CC398 isolate was detected in Group A. Contact with livestock and veterinary practice seems to increase the risk of carrying MRSA-CC398, but not in food handlers. Thus, this emphasizes the need for integrated molecular epidemiology of zoonotic staphylococci.
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Li, Shanshuang, Robert Leo Skov, Xiao Han, Anders Rhod Larsen, Jesper Larsen, Marit Sørum, Mireille Wulf, Andreas Voss, Keiichi Hiramatsu, and Teruyo Ito. "Novel Types of Staphylococcal Cassette ChromosomemecElements Identified in Clonal Complex 398 Methicillin-Resistant Staphylococcus aureus Strains." Antimicrobial Agents and Chemotherapy 55, no. 6 (March 21, 2011): 3046–50. http://dx.doi.org/10.1128/aac.01475-10.

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ABSTRACTThe structures of staphylococcal cassette chromosomemec(SCCmec) elements carried by 31 clonal complex 398 (CC398) methicillin-resistantStaphylococcus aureus(MRSA) strains isolated from the participants at a conference were analyzed. The SCCmecs were classified into novel types, namely, IX, X, V(5C2&5) subtype c, and IVa. Type V(5C2&5) subtype c, IX, and X SCCmecs carried genes conferring resistance to metals. The structures of SCCmecs from CC398 strains were distinct from those normally found in humans, adding to the evidence that humans are not the original host for CC398.
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11

Alkuraythi, Dalal M., Manal M. Alkhulaifi, Abdulwahab Z. Binjomah, Mohammed Alarwi, Hind M. Aldakhil, Mohammed I. Mujallad, Saleh Ali Alharbi, et al. "Clonal Flux and Spread of Staphylococcus aureus Isolated from Meat and Its Genetic Relatedness to Staphylococcus aureus Isolated from Patients in Saudi Arabia." Microorganisms 11, no. 12 (December 6, 2023): 2926. http://dx.doi.org/10.3390/microorganisms11122926.

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In this study, we investigated both meat-derived and methicillin-resistant Staphylococcus aureus (MRSA), exploring their genetic relatedness to patient-derived MRSA isolates in Saudi Arabia. We collected 250 meat samples and identified 53 S. aureus isolates, with 79% being methicillin-sensitive Staphylococcus aureus (MSSA) and 21% being MRSA. Moreover, we included 80 clinically confirmed patient-derived MRSA isolates. We identified the most common S. aureus clone in both patients and retail meat. In meat, ST6 and ST97 were the most common clones in 55% of the MRSA isolates, and ST1153 and ST672 were the most common in 21% and 17% of the MSSA isolates. In patients, ST5 and ST6 were the predominant clones in 46% of the S. aureus isolates. CC5/ST5-SCCmecVc-t311 and CC361/ST672-SCCmecV-t3841 were common MRSA clones in both meat and patients. CC97 and CC361 clones were the second most prevalent S. aureus clones in meat and were relatively common in patients. Furthermore, we sequenced and characterized novel S. aureus strains ST8109, ST8110, and ST8111. The genomic similarities between meat- and patient-derived S. aureus isolates suggest that retail meat might be a reservoir for S.aureus and MRSA transmission. Therefore, a structured One Health approach is recommended for S. aureus dissemination, genetic characterization, antibiotic resistance, and impact on human health.
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12

VANDERHAEGHEN, W., K. HERMANS, F. HAESEBROUCK, and P. BUTAYE. "Methicillin-resistantStaphylococcus aureus(MRSA) in food production animals." Epidemiology and Infection 138, no. 5 (February 2, 2010): 606–25. http://dx.doi.org/10.1017/s0950268809991567.

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SUMMARYUntil recently, reports on methicillin-resistantStaphylococcus aureus(MRSA) in food production animals were mainly limited to occasional detections in dairy cattle mastitis. However, since 2005 a MRSA clone, CC398, has been reported colonizing pigs, veal calves and broiler chickens and infecting dairy cows. Many aspects of its prevalence in pigs remain unclear. In other livestock, colonizing capacity and reservoir status still require elucidation. MRSA CC398 has also been detected in meat, but, as for other MRSA, the risk this poses is somewhat unclear. Currently, the most worrying aspect of MRSA CC398 appears to be its capacity to spread to humans. This might complicate MRSA control measures in human healthcare, urging research into risk factors and transmission routes. Although infections with MRSA CC398 are much less reported than carriage, more investigation into its pathogenic potential is required. Moreover, the origin and evolution of this clone remain unknown.
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Kizerwetter-Świda, Magdalena, Dorota Chrobak-Chmiel, Magdalena Rzewuska, Joanna Pławińska-Czarnak, and Marian Binek. "Characterisation of Staphylococcus aureus isolated from meat processing plants – a preliminary study." Journal of Veterinary Research 60, no. 4 (December 1, 2016): 441–46. http://dx.doi.org/10.1515/jvetres-2016-0066.

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Abstract Introduction: Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) belonging to the clonal complex 398 (CC398) emerged recently in livestock as a new type of MRSA, which may cause zoonotic infections. This study presents data on the characterisation of S. aureus isolated from the meat processing plants. Material and Methods: S. aureus was isolated from 90 samples collected in the raw meat warehouse, from devices and surfaces of meat processing plants, and from finished meat products. The isolates were subjected to molecular analysis in order to investigate the presence of enterotoxin genes, the mecA gene, and to verify whether they belong to the clonal complex 398. The genetic relatedness of the isolates was determined using pulsed-field electrophoresis. Likewise, antimicrobial susceptibility was tested. Results: From 21 S. aureus strains isolated, five belonged to the CC398, two of which were recognised as MRSA and three as methicillin-sensitive Staphylococcus aureus (MSSA). The most prevalent enterotoxin genes were seg and sei. Two MRSA CC398 isolates, three MSSA CC398, and one MSSA were classified as multidrug-resistant. Conclusion: The first isolation of MSSA CC398 from beef in Poland indicates contamination of beef by strains belonging to this clonal complex. The occurrence of multidrug-resistant enterotoxigenic S. aureus isolates in the finished meat products constitutes a potential risk for the consumers.
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van der Mee-Marquet, Nathalie, Sandra Dos Santos, Seydina M. Diene, Isabelle Duflot, Laurent Mereghetti, Anne-Sophie Valentin, and Patrice François. "Strong Biofilm Formation and Low Cloxacillin Susceptibility in Biofilm-Growing CC398 Staphylococcus aureus Responsible for Bacteremia in French Intensive Care Units, 2021." Microorganisms 10, no. 9 (September 16, 2022): 1857. http://dx.doi.org/10.3390/microorganisms10091857.

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A prospective 3-month study carried out in 267 ICUs revealed an S. aureus nosocomial bacteremia in one admitted patient out of 110 in adult and pediatric sectors, and in one out of 230 newborns; 242 S. aureus bacteremias occurred during the study, including 7.9% MRSA-bacteremias. In one ICU out of ten, the molecular characteristics, antimicrobial susceptibility profiles and biofilm production of the strains responsible for S. aureus bacteremia were studied. Of the 53 strains studied, 9.4% were MRSA and 52.8% were resistant to erythromycin. MLST showed the predominance of CC398 (37.7% of the strains) followed by CC8 (17.0%), CC45 (13.2%) and CC30 (9.4%). The lukF/S genes were absent from our isolates and tst-1 was found in 9.4% of the strains. Under static conditions and without exposure to glucose, biofilm production was rare (9.4% of the strains, without any CC398). The percentage increased to 62.3% for strains grown in broth supplemented with 1% glucose (including 7 out of 9 CC8 and 17 out of the 20 CC398). Further study of the CC398, including whole genome sequencing, revealed (1) highly frequent patient death within seven days after CC398 bacteremia diagnosis (47.4%), (2) 95.0% of the strains producing biofilm when exposed to sub-inhibitory concentrations of cloxacillin, (3) a stronger biofilm production following exposure to cloxacillin than that observed in broth supplemented with glucose only (p < 0.001), (4) a high minimum biofilm eradication concentration of cloxacillin (128 mg/L) indicating a low cloxacillin susceptibility of biofilm-growing CC398, (5) 95.0% of the strains carrying a ϕSa-3 like prophage and its particular evasion cluster (i.e., yielding chp and scin genes), and (6) 30.0% of the strains carrying a ϕMR11-like prophage and yielding a higher ability to produce biofilm. Our results provide evidence that active surveillance is required to avoid spreading of this virulent staphylococcal clone.
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Cavaco, L. M., H. Hasman, M. Stegger, P. S. Andersen, R. Skov, A. C. Fluit, T. Ito, and F. M. Aarestrup. "Cloning and Occurrence of czrC, a Gene Conferring Cadmium and Zinc Resistance in Methicillin-Resistant Staphylococcus aureus CC398 Isolates." Antimicrobial Agents and Chemotherapy 54, no. 9 (June 28, 2010): 3605–8. http://dx.doi.org/10.1128/aac.00058-10.

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ABSTRACT We recently reported a phenotypic association between reduced susceptibility to zinc and methicillin resistance in Staphylococcus aureus CC398 isolates from Danish swine (F. M. Aarestrup, L. M. Cavaco, and H. Hasman, Vet. Microbiol. 142:455-457, 2009). The aim of this study was to identify the genetic determinant causing zinc resistance in CC398 and examine its prevalence in isolates of animal and human origin. Based on the sequence of the staphylococcal cassette chromosome mec (SCCmec) element from methicillin-resistant S. aureus (MRSA) CC398 strain SO385, a putative metal resistance gene was identified in strain 171 and cloned in S. aureus RN4220. Furthermore, 81 MRSA and 48 methicillin-susceptible S. aureus (MSSA) strains, isolated from pigs (31 and 28) and from humans (50 and 20) in Denmark, were tested for susceptibility to zinc chloride and for the presence of a putative resistance determinant, czrC, by PCR. The cloning of czrC confirmed that the zinc chloride and cadmium acetate MICs for isogenic constructs carrying this gene were increased compared to those for S. aureus RN4220. No difference in susceptibility to sodium arsenate, copper sulfate, or silver nitrate was observed. Seventy-four percent (n = 23) of the animal isolates and 48% (n = 24) of the human MRSA isolates of CC398 were resistant to zinc chloride and positive for czrC. All 48 MSSA strains from both human and pig origins were found to be susceptible to zinc chloride and negative for czrC. Our findings showed that czrC is encoding zinc and cadmium resistance in CC398 MRSA isolates, and that it is widespread both in humans and animals. Thus, resistance to heavy metals such as zinc and cadmium may play a role in the coselection of methicillin resistance in S. aureus.
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ESPINOSA-GONGORA, C., J. LARSEN, A. MOODLEY, J. P. NIELSEN, R. L. SKOV, M. ANDREASEN, and L. GUARDABASSI. "Farm-specific lineages of methicillin-resistant Staphylococcus aureus clonal complex 398 in Danish pig farms." Epidemiology and Infection 140, no. 10 (November 25, 2011): 1794–99. http://dx.doi.org/10.1017/s0950268811002391.

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SUMMARYThe objective of this study was to investigate the genetic diversity of methicillin-resistant Staphylococcus aureus (MRSA) clonal complex (CC) 398 using pulsed-field gel electrophoresis (PFGE). Dust and pigs at five age groups were sampled in six Danish MRSA-positive pig farms. MRSA CC398 was isolated from 284 of the 391 samples tested, including 230 (74%) animal and 54 (68%) environmental samples. PFGE analysis of a subset of 48 isolates, including the six strains previously isolated from farm workers, revealed the existence of farm-specific pulsotypes. With a single exception, human, environmental and porcine isolates originating from the same farm clustered together in the PFGE cluster analysis, indicating that spread of MRSA CC398 in Danish pig farms is mainly due to clonal dissemination of farm-specific lineages that can be discriminated by PFGE. This finding has important implications for planning future epidemiological studies investigating the spread of CC398 in pig farming.
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Krupa, Paweł, Jarosław Bystroń, Magdalena Podkowik, Joanna Empel, Aneta Mroczkowska, and Jacek Bania. "Population Structure and Oxacillin Resistance ofStaphylococcus aureusfrom Pigs and Pork Meat in South-West of Poland." BioMed Research International 2015 (2015): 1–9. http://dx.doi.org/10.1155/2015/141475.

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The genotypes and oxacillin resistance of 420S. aureusisolates from pigs (n=203) and pork (n=217) were analyzed. Among 18spatypes detected inS. aureusfrom pig t011, t021, t034, t091, t318, t337, and t1334 were the most frequent. Among 30spatypes found inS. aureusisolates from pork t084, t091, t499, t4309, t12954, and t13074 were dominant. The animalS. aureusisolates were clustered into MLST clonal complexes CC7, CC9, CC15, CC30, and CC398 and meat-derived isolates to CC1, CC7, and CC15. Thirty-six MRSA were isolated exclusively from pigs. All MRSA were classified tospat011 SCCmecV. BORSA phenotype was found in 14%S. aureusisolates from pigs and 10% isolates from pork meat.spat034 dominated among BORSA from pigs and t091 among meat-derived BORSA. This is the first report onspatypes and oxacillin resistance ofS. aureusstrains from pigs and pork meat in Poland. BesidesS. aureusCC9, CC30, and CC398 known to be distributed in pigs, the occurrence of genotype belonging to CC7 in this species has been reported for the first time. To our knowledge it is also the first report concerning CC398 BORSA isolates from pigs and pork meat.
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Oppliger, Anne, Philippe Moreillon, Nicole Charrière, Marlyse Giddey, Delphine Morisset, and Olga Sakwinska. "Antimicrobial Resistance of Staphylococcus aureus Strains Acquired by Pig Farmers from Pigs." Applied and Environmental Microbiology 78, no. 22 (September 7, 2012): 8010–14. http://dx.doi.org/10.1128/aem.01902-12.

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ABSTRACTCarriage of animal-associated methicillin-resistantStaphylococcus aureus(MRSA) clonal complex 398 (CC398) is common among pig farmers. This study was conducted (i) to investigate whether pig farmers are colonized with pig-specificS. aureusgenotypes other than CC398 and (ii) to survey antimicrobial resistance ofS. aureusisolates from pigs and pig farmers. Forty-eightS. aureusisolates from pig farmers and veterinarians and 130 isolates from pigs collected in Western Switzerland were genotyped byspatyping and amplified fragment length polymorphism (AFLP). Antimicrobial resistance profiles were determined for representative sample of the isolates. The data obtained earlier on healthyS. aureuscarriers without exposure to agriculture were used for comparison. The genotype composition ofS. aureusisolates from pig farmers and veterinarians was similar to isolates from pigs with predominant AFLP clusters CC398, CC9, and CC49. The resistance to tetracycline and macrolides (clarithromycin) was common among the isolates from farmers and veterinarians (52 and 21%, respectively) and similar to resistance levels in isolates from pigs (39 and 23%, respectively). This was in contrast to isolates from persons without contact with agriculture, where no (0/128) isolates were resistant to tetracycline and 3% of the isolates were resistant to clarithromycin. MRSA CC398 was isolated from pigs (n= 11) and pig farmers (n= 5). These data imply that zoonotic transmission of multidrug-resistantS. aureusfrom pigs to farmers is frequent, and well-known MRSA transmission merely represents the tip of the iceberg for this phenomenon. We speculate that the relatively low frequency of MRSA isolation is related to lower antimicrobial use in Switzerland compared to, for example, the Netherlands.
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Smith, Tara C., and Shylo E. Wardyn. "Human Infections with Staphylococcus aureus CC398." Current Environmental Health Reports 2, no. 1 (January 8, 2015): 41–51. http://dx.doi.org/10.1007/s40572-014-0034-8.

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Ceballos, Sara, Carmen Lozano, Carmen Aspiroz, Laura Ruiz-Ripa, Paula Eguizábal, Allelen Campaña-Burguet, Emilia Cercenado, et al. "Beyond CC398: Characterisation of Other Tetracycline and Methicillin-Resistant Staphylococcus aureus Genetic Lineages Circulating in Spanish Hospitals." Pathogens 11, no. 3 (March 1, 2022): 307. http://dx.doi.org/10.3390/pathogens11030307.

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Tetracycline resistance (TetR) has been evidenced as a good phenotypic marker for detection of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) isolates of the clonal complex CC398. The aim of this study was to characterise a collection of 95 TetR-MRSA isolates, not belonging to the lineage CC398, that were obtained in a previous multicentre study, to detect other MRSA clonal complexes that could be associated with this phenotypic TetR marker. The TetR-MRSA isolates were recovered from 20 Spanish hospitals during 2016 and they were characterised to determine their antimicrobial resistance and virulence phenotypes/genotypes as well as the presence of the immune evasion cluster (IEC). A high proportion of isolates belonging to the CC1 lineage (46%) were observed, as well as to the CC5, CC8 and CC45 lineages (11% each one). Thirty-two different spa-types were identified, being predominantly CC1-t127 (40%) and CC45-t1081 (11%). The IEC system (with the gene scn as marker) was present in 73% of isolates and 16% produced the Panton Valentine leucocidin (PVL). A high proportion of MRSA-CC1 isolates were scn-negative (38.6%) and 52.9% of them were blaZ-negative. A multidrug resistance (MDR) phenotype was identified in 86% of MRSA isolates. The knowledge of other TetR-MRSA genetic lineages, in addition to CC398, is highly relevant, since most of them were MDR and some of them presented important virulence factors. Strains potentially associated with livestock (as the subpopulation CC1-t127-scn-negative) or with humans (as the CC45 lineage or the subpopulation CC1-scn-positive) have been found in this study. The use of tetracycline-resistance for detection, not only of CC398 but also of other LA-MRSA lineages should be tracked in the future.
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Kizerwetter-Świda, Magdalena, and Joanna Pławińska-Czarnak. "Livestock-associated strains of methicillin resistant Staphylococcus aureus (LA-MRSA) – the current state of knowledge." Medycyna Weterynaryjna 73, no. 2 (2017): 92–98. http://dx.doi.org/10.21521/mw.5639.

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A methicillin-resistant Staphylococcus aureus (MRSA) referred to as livestock-associated (LA-MRSA) has recently emerged in farm animals, particularly in pigs. Strains of this MRSA variant from Europe and North America mostly belong to clonal complex (CC) 398. Generally LA-MRSA cause asymptomatic colonization among pigs, but also in veal calves, broiler chickens, turkeys, and horses. People in contact with livestock animals are at high risk of asymptomatic colonization or infection with these bacteria. In previous years, the impact of LA-MRSA on human health was considered small. However, LA-MRSA has become more prevalent among people without direct livestock contact, especially in areas with a high density of pig production. As a result of horizontal gene transfer S. aureus CC398 strains are constantly evolving. The adaptive power of S. aureus to new hosts and acquisition of resistance to antibiotics may cause the emergence of new, more virulent clones. LA-MRSA has evolved from human-adapted methicillin-susceptible S. aureus CC398, which was proved by comparative genome analysis. The adaptation to livestock was associated with several genetic changes. The most worrying aspect of MRSA CC398 seems to be its ability to spread to humans. For this reason, continuous surveillance of further genetic changes is recommended.
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Chueahiran, Surawit, Jitrapa Yindee, Pongthai Boonkham, Nipattra Suanpairintr, and Pattrarat Chanchaithong. "Methicillin-Resistant Staphylococcus aureus Clonal Complex 398 as a Major MRSA Lineage in Dogs and Cats in Thailand." Antibiotics 10, no. 3 (February 28, 2021): 243. http://dx.doi.org/10.3390/antibiotics10030243.

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The aim of this study was to present molecular and antimicrobial resistance characteristics of methicillin-resistant Staphylococcus aureus (MRSA) clonal complex (CC) 398 isolated from diseased dogs and cats in Thailand. A total of 20 MRSA isolates of 134 Staphylococcus aureus isolated from canine and feline clinical samples during 2017–2020 were CC398, consisting of sequence type (ST) 398 (18 isolates), ST5926 (1 isolate), and ST6563 (1 isolate) by multilocus sequence typing. spa t034 and staphylococcal cassette chromosome mec (SCCmec) V were predominantly associated with ST398. Intraclonal differentiation was present by additional spa (t1255, t4653), non-detectable spa, composite SCCmec with a hybrid of ccrA1B1+ccrC and class A mec complex, and DNA fingerprints by pulsed-field gel electrophoresis. The isolates essentially carried antimicrobial resistance genes, mediating multiple resistance to β-lactams (mecA, blaZ), tetracyclines [tet(M)], aminoglycosides [aac(6′)-Ie-aph(2′)-Ia], and trimethoprim (dfr). Livestock-associated MRSA ST398 resistance genes including lnu(B), lsa(E), spw, fexA, and tet(L) were heterogeneously found and lost in subpopulation, with the absence or presence of additional erm(A), erm(B), and ileS2 genes that corresponded to resistance phenotypes. As only a single CC398 was detected with the presence of intraclonal variation, CC398 seems to be the successful MRSA clone colonizing in small animals as a pet-associated MRSA in Thailand.
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Narongpun, Pawarut, Pattrarat Chanchaithong, Junya Yamagishi, Jeewan Thapa, Chie Nakajima, and Yasuhiko Suzuki. "Whole-Genome Investigation of Zoonotic Transmission of Livestock-Associated Methicillin-Resistant Staphylococcus aureus Clonal Complex 398 Isolated from Pigs and Humans in Thailand." Antibiotics 12, no. 12 (December 16, 2023): 1745. http://dx.doi.org/10.3390/antibiotics12121745.

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Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) has been widespread globally in pigs and humans for decades. Nasal colonization of LA-MRSA is regarded as an occupational hazard to people who are regularly involved in livestock production. Our previous study suggested pig-to-human transmission caused by LA-MRSA clonal complex (CC) 398, using traditional molecular typing methods. Instead, this study aimed to investigate the zoonotic transmission of LA-MRSA CC398 using whole genome sequencing (WGS) technologies. A total of 63 LA-MRSA isolates were identified and characterized in Thailand. Further, the 16 representatives of LA-MRSA CC9 and CC398, including porcine and worker isolates, were subjected to WGS on the Illumina Miseq platform. Core-genome single nucleotide polymorphism (SNP)-based analyses verify the zoonotic transmission caused by LA-MRSA CC398 in two farms. WGS-based characterization suggests the emergence of a novel staphylococcal cassette chromosome (SCC) mec type, consisting of multiple cassette chromosome recombinase (ccr) gene complexes via genetic recombination. Additionally, the WGS analyses revealed putative multi-resistant plasmids and several cross-resistance genes, conferring resistance against drugs of last resort used in humans such as quinupristin/dalfopristin and linezolid. Significantly, LA-MRSA isolates, in this study, harbored multiple virulence genes that may become a serious threat to an immunosuppressive population, particularly for persons who are in close contact with LA-MRSA carriers.
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Broens, E. M., E. A. M. Graat, P. J. van der Wolf, A. W. van de Giessen, E. van Duijkeren, J. A. Wagenaar, A. van Nes, D. J. Mevius, and M. C. M. de Jong. "MRSA CC398 in the pig production chain." Preventive Veterinary Medicine 98, no. 2-3 (February 2011): 182–89. http://dx.doi.org/10.1016/j.prevetmed.2010.10.010.

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Argudín, M. Angeles, A. Deplano, S. Vandendriessche, M. Dodémont, C. Nonhoff, O. Denis, and S. Roisin. "CC398 Staphylococcus aureus subpopulations in Belgian patients." European Journal of Clinical Microbiology & Infectious Diseases 37, no. 5 (February 15, 2018): 911–16. http://dx.doi.org/10.1007/s10096-018-3205-y.

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26

Feßler, Andrea T., Kristina Kadlec, Melanie Hassel, Tomasz Hauschild, Christopher Eidam, Ralf Ehricht, Stefan Monecke, and Stefan Schwarz. "Characterization of Methicillin-Resistant Staphylococcus aureus Isolates from Food and Food Products of Poultry Origin in Germany." Applied and Environmental Microbiology 77, no. 20 (July 1, 2011): 7151–57. http://dx.doi.org/10.1128/aem.00561-11.

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ABSTRACTDuring a survey of fresh chicken and turkey meat as well as chicken and turkey meat products for the presence of methicillin-resistantStaphylococcus aureus(MRSA) isolates in Germany, 32 (37.2%) of 86 samples were MRSA positive. Twenty-eight of these MRSA isolates belonged to clonal complex 398 (CC398), which is widespread among food-producing animals. These CC398 isolates carried SCCmecelements of type IV or V and exhibitedspatype t011, t034, t899, t2346 or t6574 and either the knowndrutypes dt2b, dt6j, dt10a, dt10q, dt11a, dt11v, and dt11ab or the noveldrutypes dt6m, dt10as, and dt10at. In addition, two MRSA sequence type 9 (ST9) isolates with a type IV SCCmeccassette,spatype t1430, anddrutype dt10a as well as single MRSA ST5 and ST1791 isolates with a type III SCCmeccassette,spatype t002, anddrutype dt9v were identified. All but two isolates were classified as multiresistant. A wide variety of resistance phenotypes and genotypes were detected. All isolates were negative for the major virulence factors, such as Panton-Valentine leukocidin, toxic shock syndrome toxin 1, or exfoliative toxins. In contrast to the MRSA CC398 isolates, the four ST9, ST5, or ST1791 isolates harbored theegcgene cluster for enterotoxin G, I, M, N, O, and U genes. Although the relevance of contamination of fresh poultry meat or poultry products with MRSA is currently unclear, the presence of multiresistant and, in part, enterotoxigenic MRSA emphasizes the need for further studies to elucidate possible health hazards for consumers.
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Abdullahi, Idris Nasir, Rosa Fernández-Fernández, Guillermo Juárez-Fernández, Sandra Martínez-Álvarez, Paula Eguizábal, Myriam Zarazaga, Carmen Lozano, and Carmen Torres. "Wild Animals Are Reservoirs and Sentinels of Staphylococcus aureus and MRSA Clones: A Problem with “One Health” Concern." Antibiotics 10, no. 12 (December 20, 2021): 1556. http://dx.doi.org/10.3390/antibiotics10121556.

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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.
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28

D'Avila, DO, and C. Viana. Critical Care 9, Suppl 2 (2005): P74. http://dx.doi.org/10.1186/cc3618.

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29

Boswihi, Samar S., Wadha A. Alfouzan, and Edet E. Udo. "Genomic profiling of methicillin-sensitive Staphylococcus aureus (MSSA) isolates in Kuwait hospitals." Frontiers in Microbiology 15 (July 17, 2024). http://dx.doi.org/10.3389/fmicb.2024.1361217.

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BackgroundStaphylococcus aureus is an important pathogen that causes mild to invasive infections in hospitals and the community. Although methicillin-susceptible Staphylococcus aureus (MSSA) isolates continue to cause different infections, there is no data on the genetic backgrounds of the MSSA colonizing or causing infections in Kuwait hospitals. This study aimed to investigate MSSA isolated from patients admitted to Kuwait hospitals for antibiotic resistance and genetic backgrounds to understand their clonal composition.MethodsConsecutive MSSA isolates were collected from single patients during two surveillance periods in 2016 and 2021 in 13 public hospitals. The isolates were characterized using antibiogram, staphylococcal protein A (spa) typing, DNA microarray analysis, and multilocus sequence typing (MLST) using standard protocols.ResultsA total of 446 MSSA was cultured from different clinical samples in 2016 (n = 240) and 2021 (n = 206). All isolates were susceptible to vancomycin [minimum inhibitory concentration (MIC) ≤ 2 mg/L], teicoplanin (MIC ≤2 mg/L), linezolid (MIC ≤4 mg/L), ceftaroline (MIC ≤2 mg/L), rifampicin, and mupirocin but were resistant to erythromycin (21.3%), clindamycin (14.0%), gentamicin (3.8%), kanamycin (10.5%), fusidic acid (27.0%), tetracycline (6.9%), trimethoprim (23.1%), and ciprofloxacin (35.2%). Molecular typing identified 155 spa types, dominated by t127 (15.0%), t084 (5.4%), t3841 (5.4%), t267 (2.4%), t442 (2.2%), t091 (2.2%), t021 (2.2%), and t003 (2.2%); 31 clonal complexes (CCs); and 56 sequence types (STs). The majority of the isolates (n = 265; 59.4%) belonged to CC1 (20.6%), CC15 (10.9%), CC22 (5.1%), CC30 (7.6%), CC361 (10.1%), and CC398 (4.7%).DiscussionThe MSSA isolates belonged to diverse genetic backgrounds dominated by CC1, CC15, CC22, CC30, CC361, and CC398. The distribution of MSSA clones in 2016 and 2021 showed the stability of these clones over time. The study provides the first comprehensive data on the clonal distribution of MSSA in Kuwait hospitals.
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30

Soubrier, S., F. Saulnier, H. Hubert, P. Delour, H. Lenci, T. Onimus, S. Nseir, and A. Durocher. Critical Care 9, Suppl 1 (2005): P55. http://dx.doi.org/10.1186/cc3118.

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Möller, K., C. Stahl, and J. Guttmann. Critical Care 9, Suppl 1 (2005): P105. http://dx.doi.org/10.1186/cc3168.

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32

Grosjean, A., K. Zouaoui Boudjeltia, M. Piagnerelli, and M. Vanhaeverbeek. Critical Care 9, Suppl 1 (2005): P155. http://dx.doi.org/10.1186/cc3218.

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33

Knibel, M., C. David, and R. Hatum. Critical Care 9, Suppl 1 (2005): P255. http://dx.doi.org/10.1186/cc3318.

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34

Roberts, H., and N. Stallard. Critical Care 9, Suppl 1 (2005): P355. http://dx.doi.org/10.1186/cc3418.

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35

Hargrove, Jenny, and H. Bryant Nguyen. Critical Care 9, no. 4 (2005): 376. http://dx.doi.org/10.1186/cc3518.

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36

Vaisman, M., A. Parodi, FLB Gutierrez, R. Arkader, JLF Costa, PN Gomes, JRB Martins, and RCC Filho. Critical Care 9, Suppl 2 (2005): P64. http://dx.doi.org/10.1186/cc3608.

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37

Oliveira, GMMO, PH Godoy, MR Pantoja, RR Luiz, CE Figueiredo, RR Casemiro, J. Regalla, and L. Solha. Critical Care 9, Suppl 2 (2005): P66. http://dx.doi.org/10.1186/cc3610.

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38

Issa, VS, MFR Silva, LU Taniguchi, LM Cruz-Neto, and IT Velasco. Critical Care 9, Suppl 2 (2005): P67. http://dx.doi.org/10.1186/cc3611.

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39

Silva, MO, CP Amendola, S. Longui, JM Silva, EV Campos, and E. Rezende. Critical Care 9, Suppl 2 (2005): P68. http://dx.doi.org/10.1186/cc3612.

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Kopel, L., RT Carvalho, HBN Araujo, AA Fagundes, M. Ribeiro, J. Bastos, and SG Lage. Critical Care 9, Suppl 2 (2005): P69. http://dx.doi.org/10.1186/cc3613.

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41

Messeder, O., CEC Rolim, MJ Martins, MV Liberato, JM Teles, S. Agareno, and A. Farias. Critical Care 9, Suppl 2 (2005): P70. http://dx.doi.org/10.1186/cc3614.

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Silva, MFR, VS Issa, LU Tanaguchi, LM Cruz-Neto, and IT Velasco. Critical Care 9, Suppl 2 (2005): P71. http://dx.doi.org/10.1186/cc3615.

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43

Santos, MS, FL Gutierrez, JLF Costa, PN Gomes, and RC Costa-Filho. Critical Care 9, Suppl 2 (2005): P72. http://dx.doi.org/10.1186/cc3616.

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44

Rodigues, MG, F. Ruzany, DR Salgado, E. Rocha, CF Valente, E. Maccariello, and RNA Paiva. Critical Care 9, Suppl 2 (2005): P73. http://dx.doi.org/10.1186/cc3617.

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45

Mattos, MA, DG Toledo, CE Mattos, FC de Deus, MHV Assad, and BR Tura. Critical Care 9, Suppl 2 (2005): P75. http://dx.doi.org/10.1186/cc3619.

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46

Borghi-Silva, A., RG Mendes, LMM Sampaio, F. Negrini, VAP di Lorenzo, and S. Luzzi. Critical Care 9, Suppl 2 (2005): P84. http://dx.doi.org/10.1186/cc3628.

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Kurtz, PMP, CHE Falcão, MD Magalhães, D. Terrana, DA Prado, and PCP Souza. Critical Care 9, Suppl 2 (2005): P94. http://dx.doi.org/10.1186/cc3638.

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48

Lamblet, LCR, LR Guastelli, DF Moura, M. Alves, AC Bittencourt, APP Teixeira, and E. Knobel. "Peripherally inserted central catheter in critically ill patients." Critical Care 9, S2 (June 2005). http://dx.doi.org/10.1186/cc3648.

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49

Guimarães, HP, A. Raimondi, PHR Leal, RD Lopes, AP Resque, GK Barcelos, MB Peruzzo, FR Machado, and JLG Amaral. Critical Care 9, Suppl 2 (2005): P114. http://dx.doi.org/10.1186/cc3658.

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Oliveira, MC, and A. Réa-Neto. Critical Care 9, Suppl 2 (2005): P124. http://dx.doi.org/10.1186/cc3668.

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