Detection of the main multiresistant microorganisms in the environment of a teaching veterinary hospital in Brazil / Detecção dos principais microrganismos multirresistentes no ambiente de um hospital veterinário de ensino no Brasil

Contamination of the veterinary hospital environment with multiresistant pathogens endangers not only hospitalized animals, but also the workplace safety of veterinarians and nurses, animal guardians and, when in case of a teaching hospital, veterinary students. The objective of this study was to map the main points of bacterial contamination of a veterinary teaching hospital in Brazil to identify multiresistant microorganisms and their antimicrobial resistance genes. Samples were collected from 39 different locations of a veterinary school hospital which comprised a pool according to each hospital environment. In certain environments, more than one pool has been collected. All samples were collected in quadruplicates for the selective isolation of the main multiresistant microorganisms: methicillin-resistant Staphylococcus (MRS), vancomycin resistant Enterococcus (VRE), cephalosporinases and/or extended-spectrum beta-lactamase-producing Gram-negative bacteria (ESBL) and Carbapenemase-producing (CP). After isolation and identification of isolates, multiplex-PCR reactions were performed to detect the main genes for each microorganism and antimicrobial susceptibility tests with the main antibiotics used for each bacterial group according to CLSI. Of the 39 veterinary teaching hospital sites collected, all (100%) had at least one of the microorganisms surveyed, and 17.95% (n=7) of the sites were able to isolate the four pathogens. From the 94 pools collected, it was possible to isolate MRS in 81.91% (n=77), VRE in 12.77% (n=12), cephalosporinases and/or ESBL in 62.77% (n=59) and CP in 24.47%. (n=23). Regarding MRS, the mecA gene was detected in all isolates. All isolated VREs were identified as Enterococcus faecalis and presented the vanA gene. Regarding cephalosporinases and/or ESBL, 89.83% (n=53) of the isolates presented the blaTEM gene, 57.63% (n=34) the blaOXA-1 gene, 37.29% (n=22) blaCTX-M gene from some group (1, 2, 9 ou 8/25) and 20.34% (n=12) the blaSHV gene. It was possible to identify the main microorganisms responsible for causing nosocomial infections in humans (VRE, MRS, ESBL and CP) in the veterinary hospital environment, suggesting a source of infection for professionals and students of veterinary medicine, placing a high risk for public health.(AU)

A contaminação do ambiente hospitalar veterinário com patógenos multirresistentes coloca em perigo não apenas os animais hospitalizados, mas também a segurança no local de trabalho de veterinários e enfermeiros, responsáveis por animais e, quando se tratar de um hospital de ensino, estudantes de veterinária. O objetivo deste estudo foi mapear os principais pontos de contaminação bacteriana de um hospital veterinário de ensino no Brasil, identificando microorganismos multirresistentes e seus genes de resistência antimicrobiana. As amostras foram coletadas em 39 locais diferentes de um hospital de escola veterinária, que compreendia um pool de acordo com o ambiente de cada hospital. Em certos ambientes, mais de um pool foi coletado. Todas as amostras foram coletadas em quadruplicados para o isolamento seletivo dos principais microorganismos multirresistentes: Staphylococcus resistente à meticilina (MRS), Enterococcus resistente à vancomicina (VRE), bactérias Gram-negativas produtoras de cefalosporinases e/ou beta-lactamase de espectro estendido (ESBL) e produtoras de carbapenemase (PC). Após o isolamento e identificação dos isolados, foram realizadas reações de PCR multiplex para detectar os principais genes de cada microorganismo e testes de susceptibilidade a antimicrobianos com os principais antibióticos utilizados para cada grupo bacteriano de acordo com o CLSI. Dos 39 locais do VCH coletados, todos (100%) possuíam pelo menos um dos microrganismos pesquisados e 17,95% (n=7) dos locais foram capazes de isolar os quatro patógenos. Dos 94 pools coletados, foi possível isolar MRS em 81,91% (n=77), VRE em 12,77% (n=12), ESBL em 62,77% (n=59) e carbapenemases em 24,47% (n=23). Em relação ao MRS, o gene mecA foi detectado em todos os isolados. Todos os VREs isolados foram identificados como Enterococcus faecalis e apresentaram o gene vanA. Em relação às cefalosporinases e/ou ESBL, 89,83% (n=53) dos isolados apresentaram o gene blaTEM, 57,63% (n=34) o gene blaOXA-1, 37,29% (n=22) o gene blaCTX-M de algum grupo e 20,34% (n=12) o gene blaSHV. Foi possível identificar os principais microrganismos responsáveis por causar infecções nosocomiais em humanos (VRE, MRS, ESBL e CP) no ambiente hospitalar veterinário, sugerindo uma fonte de infecção para profissionais e estudantes de medicina veterinária, colocando alto risco para a saúde pública.(AU)

Anti-Staphylococcus aureus Methicillin-Resistant (MRSA) activity of a novel 3-Chalcogenyl Indole / Atividade Anti-Staphylococcus aureus Meticilina Resistente (MRSA) de um novo composto 3-Calcogenil Indol

Objective: the development of new drugs against Methicillin-resistant Staphylococcus aureus is a priority to the World Health Organization. So, the objective of this study was to evaluate the antibacterial activity and toxicity of 5-bromo-3-((4-methoxyphenyl) sulfenyl)-1H-indole (3b) against MRSA. Methods: minimum inhibitory concentration (MIC) of 3b was determined against S. aureus ATCC 29213 and 43 clinical isolates. The time-kill assay was performed for 9 isolates. Analysis of variance followed by the post hoc Bonferroni test was used for the statistical tests. Results and conclusions: the MIC50 and MIC90 of 3b were 4 µg.mL-1 and 16 µg.mL-1 respectively. In time-kill assay, the 3b showed bactericidal activity to all evaluated isolates at concentrations of 1xMIC and 2xMIC and the re-growth effect was not observed. About the toxicity tests, 3b has not presented cytotoxicity, mutagenicity, or allergenicity. 3b had particularly good activity against MRSA demonstrating high potential for the development of new antimicrobials products.

Objetivo: o desenvolvimento de novos antimicrobianos contra Staphylococcus aureus resistentes à meticilina (MRSA) é uma prioridade para a Organização Mundial da Saúde. Então, o objetivo desse estudo foi avaliar a atividade antibacteriana e a toxicidade do 5-bromo-3-((4-metoxifenil) sulfenil)-1H-indol (3b) contra MRSA. Métodos: a concentração inibitória minima de 3b foi determinada contra S. aureus ATCC 29213 e 43 isolados clínicos. O ensaio de curva de morte foi realizado para nove isolados. Análise de variância seguida pelo teste post hoc Bonferroni foi usada para testes estatísticos. Resultados e conclusões: a MIC50 e MIC90 do 3b foi 4 µg.mL-1 e 16 µg.mL-1, respectivamente. No ensaio de curva de morte, o 3b demonstrou atividade bactericida contra todos os isolados avaliados na concentração de 1xMIC e 2xMIC e o recrescimento não foi observado. Em relação aos testes de toxicidade, 3b não apresentou citotoxicidade, mutagenicidade ou alergenicidade. 3b apresentou atividade particularmente interessante contra MRSA, demonstrando alto potencial para o desenvolvimento de novos produtos antimicrobianos.

Virulence genes profile and biofilm formation of Methicillin-resistant and Methicillin-susceptible Staphylococcus aureus isolates from diabetic foot infections

Aims@#Diabetic foot infections (DFIs) represent one of the most important risk factors for lower extremity amputation. One of the major infection agents that causes DFIs is Staphylococcus aureus. Staphylococcus aureus is an important human pathogen causing variety of clinical manifestations which can lead to invasive infections, sepsis and even death. Outcomes of antibiotic treatment of diabetic foot infections may depend not only on the antimicrobial susceptibility of the etiological agents, but also their ability to produce diverse virulence factors. This study was aimed to investigate biofilm production and the presence of various virulence genes among Methicillin-resistant Staphylococcus aureus (MRSA) and Methicillin-susceptible Staphylococcus aureus (MSSA) isolates obtained from patients with DFIs. @*Methodology and results@#A total of 48 clinical MRSA and MSSA isolates obtained from diabetic foot patients were studied for their biofilm formation and the presence of 29 known virulence genes. The biofilm formation was observed, analyzed and quantified using the microtiter plate method. Biofilm production was observed as 95.50% and 92.00% in the MRSA and MSSA isolates, respectively. Among the 29 virulence genes tested on the 48 clinical isolates, 19 virulence genes were detected. It was found that aap (62.50%), etd (54.17%), icaD (50.00%), aae (50.00%), seh (31.25%) and icaADB (22.92%) were the most prevalent genes. A total of 10 virulence genes (etb, gehD, icaB, icaC, seb, hla_haem, hld_epid, altE, fbe and sesI) were absent in all the isolates used.@*Conclusion, significance and impact of study@#Virulence genes play important role in clinical infections. Our results showed the presence rates of biofilm formation and accumulation-associated factors that are high among MRSA as well as MSSA isolates from DFIs. These results confirmed the importance of biofilm formation as regarded for DFIs.

Índice de contaminação por ORSA em superfícies de uma enfermaria de Infectologia em Vitória, ES / ORSA contamination index in surfaces of an Infectology nursing in Vitória, ES

Objetivo: Avaliar a presença de Staphylococcus aureus resistente à oxacilina (ORSA) em superfícies frequentemente tocadas pelas mãos dos pacientes e profissionais de saúde (unidades de amostragem: maçanetas de portas e grades laterais dos leitos) antes e depois de limpeza concorrente em dias de semana e no final de semana. Método: Trata-se de estudo transversal de abordagem qualitativa, realizado na enfermaria de Infectologia de um Hospital Universitário de Vitória, ES. A qualidade da desinfecção foi avaliada por meio de cultivo qualitativa de S. aureus em uma área delimitada dos locais de coleta antes e após a limpeza concorrente. Posteriormente foi avaliado por meio de teste de disco-difusão o perfil de susceptibilidade das linhagens de S. aureus frente à oxacilina. As variáveis estudadas foram presença da bactéria e perfil de susceptibilidade (antibiograma). Resultados: Foram coletadas 93 amostras, sendo 37 (39,78%) em grades de leitos e 56 (60,22%) em maçanetas com proporção de dias de semana e final de semana semelhantes. Das 93 amostras, vinte (21,51%) foram positivas para S. aureus. Destas, quatro (20%) foram identificadas como ORSA. Conclusão: A análise estatística por meio do teste de Fisher revelou que não existe associação entre a qualidade, antes e depois, da limpeza. A análise entre os dias de coleta, final de semana e dias de semana, revelou que há independência entre as variáveis, corroborando a presença de um padrão de limpeza, independente do dia da semana.

Objective: To evaluate the presence of oxacillin-resistant Staphylococcus aureus (ORSA) on surfaces frequently touched by the hands of patients and healthcare professionals (sampling units: door handles and bed side rails) before and after concurrent cleaning on weekdays and at the weekend. Methods: This is a cross-sectional study with a qualitative approach, carried out in the Infectious Diseases of a University Hospital in Vitória, ES. The quality of disinfection was assessed by means of qualitative cultivation of S. aureus in a defined area of the collection sites before and after concurrent cleaning. Subsequently, the susceptibility profile of S. aureus strains against oxacillin was evaluated by means of a disk-diffusion test. The variables studied were the presence of the bacterium and susceptibility profile (antibiogram). Results: 93 samples were collected, 37 (39.78%) in bed racks and 56 (60.22%) on door handles with a similar proportion of weekdays and weekends. Of the 93 samples, twenty (21.51%) were positive for S. aureus. Of these, four (20%) were identified as ORSA. Conclusion: Statistical analysis using Fisher's test revealed that there is no association between the quality, before and after, of cleaning. The analysis between the collection days, weekends and weekdays, revealed that there is independence between the variables, corroborating the presence of a cleaning pattern, regardless of the day of the week.

Complete genome sequence of a methicillin-resistant Staphylococcus schleiferi strain from canine otitis externa in Korea

Livestock-associated methicillin-resistant Staphylococcus aureus in Korea: antimicrobial resistance and molecular characteristics of LA-MRSA strains isolated from pigs, pig farmers, and farm environment

Identification and characterization of methicillin-resistant Staphylococcus spp. isolated from surfaces near patients in an intensive care unit of a hospital in southeastern Brazil

Abstract INTRODUCTION: Contaminated hospital environments contribute to the transmission of microorganisms associated with healthcare. Contaminated surfaces handled by patients or healthcare professionals are a source of microorganism transmission by hand. Methicillin-resistant Staphylococcus bacteria are among the main agents responsible for increasing healthcare-associated infections in Brazil and worldwide. METHODS: The objective of this study was to screen and characterize methicillin-resistant Staphylococcus spp. on surfaces near patients in an intensive care unit. Microbiological samples, collected from ten beds in an intensive care unit with five sampling sites, were inoculated into a methicillin-resistant Staphylococcus aureus chromogenic medium. MALDI-TOF and PCR analyses were used to identify the bacteria. Antimicrobial susceptibility was determined using the disk diffusion test. The presence of the mecA gene was investigated using PCR. RESULTS: We observed that 44 out of the 50 sampling sites presented grown isolates in the methicillin-resistant Staphylococcus aureus medium. The incidence of isolated microorganisms on the right side rail, left side rail, tables, infusion pump keypad, and cardiac monitor were 18.8 %, 36.7 %, 10.9 %, 2.4 %, and 31 %, respectively. The 42 isolates included in this study were identified as coagulase-negative Staphylococcus. All of these microorganisms were multidrug-resistant and mecA gene-positive. CONCLUSIONS: This study identified the presence of methicillin-resistant coagulase-negative Staphylococcus on the beds of an intensive care unit, providing evidence for the necessity of assertive actions to decrease the risk of healthcare-associated infections at the site.

Ocorrência de Staphylococcus spp. resistente à meticilina em otite externa canina / Occurrence of methicillin-resistant Staphylococcus spp. in canine external otitis

Objetivo: O presente estudo objetivou traçar o perfil de resistência dos isolados de Staphylococcus spp. resistentes à meticilina (SMR) em cães com otite externa proveniente de atendimentos ambulatoriais ou de internação. Métodos: Isolamento e identificação bacteriológica por meio de provas bioquímicas e testes de susceptibilidade a antibacterianos. Detecção do gene mecA por PCR e determinação da concentração bactericida mínima (CBM) ao gluconato de clorexidina. Resultados: Foram coletadas 140 amostras e, destas, foram isolados 122 Staphylococcus spp. (49,4%). Dos isolados bacterianos, 14 cepas (11,47%) foram SMR (oito coagulase positiva e seis coagulase negativa), apresentando o gene mecA. A concentração bactericida mínima ao gluconato de clorexidina para os Staphylococcus spp. coagulase positiva foi de 500.000 mg/mL (0,5%) e para os coagulase negativa foi de 62.500 mg/mL (0,0625%). Cinco SMR foram positivos para o teste-D. Os SMR foram 100% sensíveis à linezolida, cloranfenicol e rifampicina. Conclusão: Apesar da baixa frequência da multirresistência encontrada, existe a necessidade de monitoramento efetivo em estirpes isoladas de animais domésticos, garantindo o sucesso do tratamento e controle da resistência bacteriana em infecções otológicas caninas.

Objective: The objective of this study is to outline the resistance profile of methicillin-resistant Staphylococcus spp. (MRS) isolates from outpatient and inpatient care dogs diagnosed with otopathy. Methods: Isolation and bacteriological identification through biochemical tests and antibacterial susceptibility testing. Detection of mecA gene by PCR and determination of minimum bactericidal concentration (CBM) to chlorhexidine gluconate. Results: 140 samples were collected and from these 122 was Staphylococcus spp. (49.4%). Of all bacterial isolates 14 (11.47%) were MRS (eight coagulase positive and six coagulase negative) with mecA gene. The minimum bactericidal concentration to chlorhexidine guconate was 500,000 mg/mL (0.5%) for coagulase positive Staphylococcus spp. and 62,500 mg/mL (0.0625%) for coagulase negative. Five MRS were positive for D-test. The MRS were 100% sensitive to linezolide, chloramphenicol and rifampicin. Conclusion: Despite the low frequency of multiressitance found, there is a need for more effective monitoring in strains isolated from domestic animals to guarantee successful treatment and control of bacterial resistance in canine otological infections.

Resistencia a la meticilina y producción de biopelícula en aislamientos clínicos de Staphylococcus aureus y Staphylococcus coagulasa negativa en México / Methicillin resistance and biofilm production in clinical isolates of Staphylococcus aureus and coagulase-negative Staphylococcus in México

Resumen Introducción. Las infecciones por Staphylococcus aureus y Staphylococcus coagulasa negativa multirresistentes a los antibióticos y asociadas con la atención en salud tienen un gran impacto epidemiológico por su alta morbimortalidad; además, se han relacionado con la formación de biopelículas, lo cual también se asocia con la resistencia a los antimicrobianos. Objetivo. Determinar la resistencia a la meticilina y cuantificar la producción de biopelículas para establecer su posible relación con los aislamientos clínicos de S. aureus y Staphylococcus coagulasa negativa. Materiales y métodos. Se estudiaron 11 cepas de S. aureus y 12 de Staphylococcus coagulasa negativa. La resistencia a la meticilina se determinó con discos de cefoxitina tomando como valores de referencia los estándares del Clinical Laboratory Standards Institute (CLSI) de 2018. La producción de biopelícula se cuantificó con cristal violeta. Los genes mecA e icaADBC se identificaron mediante reacción en cadena de la polimerasa (PCR), y se hizo un análisis bivariado con la prueba de ji al cuadrado y el coeficiente V de Cramér, utilizando el programa SPSS™, versión 20.0. Resultados. Nueve cepas de S. aureus fueron resistentes a la meticilina (SARM) y dos fueron sensibles. Ocho cepas de Staphylococcus coagulasa negativa fueron resistentes y cuatro fueron sensibles. El genotipo mecA se encontró en ocho de las nueve cepas de S. aureus y en seis de las ocho de Staphylococcus coagulasa negativa resistentes a meticilina. Todas las cepas formaron biopelícula. Diez cepas de S. aureus y 11 de Staphylococcus coagulasa negativa presentaron el genotipo icaADCB. No se encontró asociación entre la resistencia a meticilina y la formación de biopelícula. Conclusiones. La cefoxitina es suficiente para determinar el fenotipo resistente a meticilina y se asoció con el genotipo mecA. Las cepas resistentes a la meticilina y poseedoras del gen mecA pueden presentar un mecanismo de resistencia alterno. Los dos grupos de cepas formadoras de biopelícula se relacionaron con la presencia del operón icaADCB. La formación de biopelícula y la resistencia a la meticilina se expresaron como características independientes en los dos grupos de cepas.

Abstract Introduction: Infections associated with health care caused by S. aureus and coagulase- negative Staphylococci multi-resistant to antibiotics cause a high epidemiological impact due to their high morbidity and mortality. Biofilm formation, which has been associated with antimicrobial resistance, can also occur. Objectives: To determine methicillin resistance and to quantify the biofilm production to establish if there is a relationship in clinical isolates of S. aureus and coagulase-negative Staphylococci. Material and methods: A total of 11 strains of S. aureus and 12 of coagulase-negative Staphylococci were studied. Methicillin resistance was determined with cefoxitin discs and the Clinical Laboratory Standards Institute (CSLI), 2018 reference values. Biofilm production was quantified by the crystal violet method. The mecA and icaADBC genes were identified by PCR. A bivariate analysis was performed with chi-square (c2) and Cramér's V statistical tests, using SPSS™, version 20.0 software. Results: Nine S. aureus strains were methicillin-resistant and two were sensitive. Eight coagulase-negative Staphylococci strains were resistant and four were sensitive. The mecA genotype was found in eight of the nine S. aureus resistant strains and six of eight resistant coagulase-negative Staphylococci. All strains formed biofilms. Ten strains of S. aureus and 11 of coagulase-negative Staphylococci presented the icaADCB genotype. No association was found between methicillin-resistance and biofilm formation. Conclusions: Cefoxitin is enough to define the resistance phenotype and is associated with the mecA genotype. All strains formed biofilms and were related to the presence of the icaADCB operon. Biofilm formation and methicillin resistance were independent features in both groups of strains.

Specification of Bacteriophage Isolated Against Clinical Methicillin-Resistant Staphylococcus Aureus

OBJECTIVES: The emergence of resistant bacteria is being increasingly reported around the world, potentially threatening millions of lives. Amongst resistant bacteria, methicillin-resistant Staphylococcus aureus (MRSA) is the most challenging to treat. This is due to emergent MRSA strains and less effective traditional antibiotic therapies to Staphylococcal infections. The use of bacteriophages (phages) against MRSA is a new, potential alternate therapy. In this study, morphology, genetic and protein structure of lytic phages against MRSA have been analysed. METHODS: Isolation of livestock and sewage bacteriophages were performed using 0.4 μm membrane filters. Plaque assays were used to determine phage quantification by double layer agar method. Pure plaques were then amplified for further characterization. Sulfate-polyacrylamide gel electrophoresis and random amplification of polymorphic DNA were run for protein evaluation, and genotyping respectively. Transmission electron microscope was also used to detect the structure and taxonomic classification of phage visually. RESULTS: Head and tail morphology of bacteriophages against MRSA were identified by transmission electron microscopy and assigned to the Siphoviridae family and the Caudovirales order. CONCLUSION: Bacteriophages are the most abundant microorganism on Earth and coexist with the bacterial population. They can destroy bacterial cells successfully and effectively. They cannot enter mammalian cells which saves the eukaryotic cells from lytic phage activity. In conclusion, phage therapy may have many potential applications in microbiology and human medicine with no side effect on eukaryotic cells.

Clinical Features of Staphylococcal Scalded Skin Syndrome Caused by Community-Associated Methicillin-Resistant Staphylococcus aureus in Changwon City, Korea, during 2006 and 2015

PURPOSE: We investigated the clinical features and epidemiology of staphylococcal scalded skin syndrome (SSSS) from year 2006 to 2015 in Changwon city, Korea. METHODS: We reviewed medical records of 69 patients diagnosed with SSSS from year 2006 to 2015. Antibiotic susceptibility testing was performed by agar dilution method. Methicillin-resistant Staphylococcus aureus (MRSA) was phenotypically identified by oxacillin susceptibility testing and genotypically confirmed by the existence of the mecA gene. RESULTS: The median age of patients was 2.0 years (range 0.2–6 years). Three (4.3%), 53 (76.8%), and 13 (18.9%) patients showed the generalized type, the intermediate type, and the abortive type, respectively. Patients occurred throughout the year, but most patients occurred between July and October. MRSA was isolated from 54 of the 60 patients regardless of the clinical types. All patients recovered without any complications. CONCLUSIONS: There was a constant occurrence of SSSS patients caused by MRSA in Changwon area during 2006 and 2015. It is needed to constantly monitor the occurrence of patients with SSSS.

Carriage of Staphylococcus schleiferi from canine otitis externa: antimicrobial resistance profiles and virulence factors associated with skin infection

The recent emergence of Staphylococcus schleiferi in dogs with otitis externa or skin and soft tissue infections has become a significant zoonotic issues. In the current study, we investigated 1) the carriage rates of S. schleiferi among major staphylococci in healthy dogs and dogs with otitis externa, 2) antibiotic susceptibility profiles of S. schleiferi, particularly methicillin resistance (MR), and 3) virulence factors associated with skin and soft tissue infections such as ability to form biofilm, resistance to cationic antimicrobial peptides (CAMPs), and carriage of staphylococcal enterotoxin genes. Among the 21 S. schleiferi isolates, 5 isolates (24%) were determined to be methicillin-resistant (MRSS). Staphylococcal cassette chromosome mec (SCCmec) typing revealed the presence of SCCmec type V in 4 MRSS isolates and type VII in one MRSS. Higher levels of antibiotic resistance, especially multidrug resistance, were observed in MRSS isolates compared to the methicillin-susceptible S. schleiferi (MSSS) isolates. In addition, MRSS isolates exhibited enhanced ability to form biofilm under static condition and all the 5 MRSS isolates carried three or more enterotoxin genes. However, there were no significant differences in resistance to CAMPs between MRSS and MSSS isolates. These findings suggest that coagulase-negative S. schleiferi is becoming more prevalent in canine otitis externa cases. Our results also highlight the presence of multidrug-resistant MRSS isolates with enhanced biofilm production and carriage of multiple enterotoxins.

Interactions Between Atopic Dermatitis and Staphylococcus aureus Infection: Clinical Implications

Staphylococcus aureus commonly colonizes the skin of atopic dermatitis (AD) patients and contributes to the development and exacerbation of AD. Multiple factors are associated with colonization of AD skin by S. aureus, including the strength of S. aureus-corneocyte adhesion, deficiency of antimicrobial peptides, decreased levels of filaggrin and filaggrin degradation products, overexpressed Th2/Th17 cytokines, microbial dysbiosis and altered lipid profiles. S. aureus colonization on AD skin causes skin barrier dysfunction through virulence factors such as superantigens (toxins), enzymes and other proteins. Furthermore, colonization of AD skin by S. aureus exacerbates AD and may contribute to microbial dysbiosis, allergen sensitization, Th2/Th17 polarization, development of atopic march and food allergy in AD patients. Skin colonization of S. aureus, particularly methicillin-resistant S. aureus (MRSA), is one of the major challenges commonly encountered in the management of AD. Bleach bath, and topical or systemic antibiotics could be used to control S. aureus infection on AD skin. However, careful use of antibiotics is required to control the occurence of MRSA. Recently, various strategies, including microbiome transplant, monoclonal antibodies against virulent toxins, vaccines and recombinant phage endolysin, have been studied to control S. aureus infection on AD skin. Further advances in our understanding of S. aureus could provide us with ways to manage S. aureus colonization more effectively in AD patients.

Delayed intraorbital infection after craniofacial bone surgery

Intraorbital infection shows a low incidence, but it might cause blindness or even death. This case is unusual in that its origin from a craniofacial bone fracture prior to infection of the maxillary sinus. A 33-year-old female patient was referred for right cheek swelling. When she visited the emergency room, we removed right cheek hematoma and bacterial examination was done. In the past, she had craniofacial bone surgical history due to a traffic accident 6 years ago. Next day, the swelling had remained with proptosis and pus was recognized in the conjunctiva. We planned an emergency operation and removed the pus which was already spread inside the orbit. And the evaluation for sinusitis was consulted to the otorhinolaryngology department simultaneously. There were Prevotella oralis and methicillin-resistant Staphylococcus epidermidis bacterial infection in the intraorbital and sinus respectively. Afterwards, the vigorous dressing was done for over a month with intravenous antibiotics. Though the intraorbital infection was resolved, blindness and extraocular movement limitation were inevitable. In conclusion, close follow up of the maxillary sinus in facial bone fracture patients is important and aggressive treatment is needed when an infection is diagnosed.

Trends in Bloodstream Infections and Antimicrobial Susceptibilities at a University Hospital in Korea Between 2007 and 2016

BACKGROUND: Blood culture is an important method for identifying infectious microorganisms and confirming that a selected antimicrobial treatment is appropriate. In this study, we investigated the annual changes in the frequencies of blood isolates and antibiotic susceptibility test (AST) results. METHODS: We created a large database comprising data on all patient-unique blood cultures obtained from January 2007 through December 2016. Blood specimens were cultured using the BD BACTEC FX system, and species identification and AST were performed using the VITEK 2 system. RESULTS: During the 10-year study period, a total of 203,651 blood culture results were collected. Of these, gram-positive cocci, gram-negative rods, and fungi were isolated in 2.15%, 0.55%, and 0.12% of the blood cultures, respectively. Escherichia coli was the most commonly isolated species (22.8%), followed by Staphylococcus epidermidis (16.8%), Klebsiella pneumoniae (8.1%), and Staphylococcus aureus (8.0%). Fungal species were isolated in 3.0% of all positive blood cultures. Candida albicans was the most commonly isolated species (1.1%), followed by Candida parapsilosis (0.6%). Methicillin resistance was seen in 55.2% of S. aureus isolates. The frequencies of vancomycin-resistant Enterococcus (VRE) and carbapenem-resistant Pseudomonas aeruginosa (CRPA) were 13.1% and 10.9%, respectively. The isolation rates of MRSA, VRE, and CRPA showed different patterns each year. CONCLUSIONS: Among the isolates, E. coli was the most common, followed by S. epidermidis and K. pneumoniae. This study represents a long-term analysis of bloodstream infections, and the results can be used to identify trends in the microorganisms isolated and their drug resistance.

Genetic Variability of Methicillin Resistant Staphylococcus Aureus Strains Isolated from Burns Patients

OBJECTIVES: Staphylococcus aureus is a nosocomial pathogen that provides a major challenge in the healthcare environment, especially in burns units where patients are particularly susceptible to infections. In this study, we sought to determine molecular types of S. aureus isolates collected from burns patients, based on staphylococcal protein A and coagulase gene polymorphisms. METHODS: Antibiotic susceptibility testing of 89 S. aureus strains isolated from burn wounds of patients was assessed using the Kirby-Bauer disk diffusion method. Strains were characterized by spa typing, coa typing, and resistance and toxin gene profiling. RESULTS: A total of 12 different spa types were identified with the majority being t790 (18%). Panton-Valentine leucocidin encoding genes were identified in spa types t044 (5.6%), t852 (2.2%) and t008 (2.2%). The most commonly detected antibiotic resistance gene was ant (4′)-Ia (60.7%). Ten different coa types were detected and the majority of the tested isolates belonged to coa III (47.2%). All the high-level mupirocin-resistant and low-level mupirocin resistant strains belonged to coa type III. CONCLUSION: The present study illustrated that despite the high frequency of coa III and spa t790 types, the genetic background of S. aureus strains in Iranian burns patients was diverse. The findings obtained are valuable in creating awareness of S. aureus infections within burns units.

Molecular Epidemiologic Study of a Methicillin-resistant Staphylococcus aureus Outbreak at a Newborn Nursery and Neonatal Intensive Care Unit

PURPOSE: This study aimed to investigate the molecular epidemiology of a methicillin-resistant Staphylococcus aureus (MRSA) outbreak at a newborn nursery and neonatal intensive care unit (NICU).METHODS: During the outbreak, from August to September 2017, MRSA isolates collected from neonates and medical staff underwent genotyping and screened for virulence factors. Antibiotic susceptibilities were tested.RESULTS: During the study period, 41 neonates were admitted at the nursery (n=27) and NICU (n=14). Of these, 7 had MRSA infections (skin infection [n=6] and sepsis [n=1]) and 4 were colonized with MRSA. Associated medical staff (n=32) were screened; three were nasal MRSA carriers. Staphylococcal chromosomal cassette mec (SCCmec) type II, sequence type (ST) 89, spa type t375 was found to be the skin infection outbreak causing strain, with multi-drug resistance including low-level mupirocin resistance. SCCmec type IVa, ST 72, and a novel spa type designated t17879, was the cause of MRSA sepsis. Many different types of MRSA were colonized on the neonates; however, SCCmec type IVa, ST 72, spa type t664 was colonized in both neonates and a NICU nurse. All MRSA isolates from colonized infants were positive for the Panton-Valentine leukocidin (PVL) toxin gene.CONCLUSIONS: The strain causing an outbreak of skin infections had multi-drug resistance. Also, MRSA colonized in the neonates were found to carry the PVL toxin gene. Because different strains are present during an outbreak, molecular epidemiologic studies are important to identify the outbreak strain and colonized strains which aid in effective control and prevention of future MRSA outbreaks.

Changing Susceptibility of Staphylococcus aureus in Children with Skin and Soft Tissue Infections: a Single Center Experience from 2010 to 2018

PURPOSE: Staphylococcus aureus is a major cause of skin and soft tissue infections (SSTIs). This study aimed to determine the temporal trends in antibiotic susceptibility of S. aureus in SSTI patients aged <19 years.METHODS: This retrospective observational study was conducted in pediatric patients with SSTI caused by community-associated S. aureus. Microbiologic and demographic data were collected, and the trends of antibiotic susceptibility results were evaluated.RESULTS: From January 2010 to December 2018, a total of 807 S. aureus isolates were included. An overall increase in susceptibility of isolates to oxacillin was noted (P<0.001), with 75.0% of isolates being oxacillin-susceptible in 2018. S. aureus remained highly susceptible to trimethoprim/sulfamethoxazole and tetracycline, with 97.6% and 95.2% isolate susceptibility in 2018, respectively. Isolates from younger children aged 1 to 5 years had a significantly lower rate of susceptibility to oxacillin than older children aged 6 to 18 years (53.4% vs. 75.0%, P<0.001).CONCLUSIONS: The proportion of methicillin-resistant S. aureus isolates appears to decrease in pediatric patients with community-associated SSTI caused by S. aureus. Clinicians should be aware of regional susceptibility patterns when choosing empirical regimens.

Chronic Recurrent Methicillin Resistant Staphylococcus Hemolyticus Endophthalmitis after Cataract Surgery

PURPOSE: To report a case of recurrent endophthalmitis due to methicillin resistant Staphylococcus hemolyticus after phacoemulsification and posterior chamber intraocular lens (IOL) implantation. CASE SUMMARY: A 76-year-old female visited our outpatient clinic with decreased vision 40 days after uncomplicated cataract surgery in her right eye. At the visit, anterior chamber inflammation and cloudy fluid between the posterior capsule and IOL were observed. Uveitis due to residual cortex of lens or capsular block syndrome was suspected, so YAG laser capsulotomy and subconjunctival injection of dexamethasone were performed. Two days later, hypopyon and vitreous opacity were seen. The patient underwent an emergency vitrectomy and intravitreal antibiotic injection with suspicion of bacterial endophthalmitis. The culture was negative. Twenty days after the vitrectomy, anterior chamber inflammation and vitreous opacity developed. The recurrence of endophthalmitis was suspected due to infection by bacteria in the surrounding tissue of the IOL, so the patient underwent an IOL and lens capsule removal with intravitreal antibiotic injection. At this time, the culture revealed methicillin resistant staphylococcus hemolyticus. Systemic and topical vancomycin was then administered, resulting in decreased inflammation. Twenty days after the IOL removal, decreased vision, anterior chamber inflammation, and vitreous opacity developed. Endophthalmitis was decreased by intravitreal antibiotic injection and topical antibiotic treatment. CONCLUSIONS: Methicillin resistant staphylococcus hemolyticus should be considered in the differential diagnosis of chronic recurrent endophthalmitis after cataract surgery.

Molecular characterization of methicillin-resistant Staphylococcus aureus isolates from a hospital in Ghana

Background: Methicillin-resistant Staphylococcus aureus (MRSA) are a major cause of hospital- and community-acquired infection. They can colonize humans and cause a wide range of infections including pneumonia, endocarditis and bacteraemia. We investigated the molecular mechanism of resistance and virulence of MRSA isolates from a teaching hospital in Ghana. Methodology: A total of 91 S. aureus isolates constituted the initial bacterial sample. Identification of S. aureus was confirmed by the VITEK 2 system. The cefoxitin screen test was used to detect MRSA and antibiotic susceptibility was determined using the VITEK 2 system. The resistance (mecA, blaZ, aac-aph, ermC, and tetK) and virulence (lukS/F-PV, hla, hld and eta) genes were amplified by polymerase chain reaction (PCR) and positive samples subjected to DNA sequencing. Pulsed field gel electrophoresis (PFGE) was used to ascertain the relatedness of the isolates. Results: Fifty-eight of 91 (63.7%) isolates were putatively methicillin resistant by the phenotypic cefoxitin screen test and oxacillin MICs. However, 43 (47%) of the isolates were genotypically confirmed as MRSA based on PCR detection of the mecA gene. Furthermore, 37.9% of isolates displayed resistance to tetracycline, 19% to trimethoprim-sulphamethoxazole, 15.5% to clindamycin, 12.1% to gentamicin, 13.8% to ciprofloxacin and erythromycin, 6.9% to moxifloxacin and 7.0% to rifampicin. None of the isolates was positive for inducible clindamycin resistance. The prevalence of resistance (mecA, blaZ, aac(6')-aph(2''), tetK, and ermC) and virulence (hla and lukS/F-PV) genes respectively were 74%, 33%, 22%, 19%, 3%, 5% and 3%, with isolates organized in two highly related clades. Conclusion: Results indicate a fairly high occurrence of MRSA, which can complicate the effective therapy of S. aureus infections, necessitating surveillance and stringent infection control programmes to forestall its spread