Detection of multidrug-resistant bacteria in the nasal cavities and evaluation of sinus disorders in patients undergoing Le Fort I osteotomy.

INTRODUCTION: Orthognathic surgery can lead to sinus alterations, including sinusitis, attributed to the exposure of maxillary sinuses during Le Fort I osteotomy. Furthermore, being a hospital-based procedure, there is potential risk of complications arising from bacteria prevalent in such environments. This study evaluated maxillary sinusitis occurrence and the presence of multidrug-resistant bacteria in the nasal cavity before and after orthognathic surgery. METHODS: Ten patients with dentofacial deformities underwent Le Fort I osteotomy. Clinical evaluations using SNOT-22 questionnaire were performed, and nasal cavity samples were collected pre-surgery and 3-6 months post-surgery to quantify total mesophilic bacteria and detect Staphylococcus aureus, Acinetobacter baumannii, and Klebsiella pneumoniae. Cone Beam Computed Tomography (CBCT) was performed pre- and post-operatively, and the results were evaluated using the Lund-Mackay system. This study was registered and approved by the Research Ethics Committee of PUCRS (No. 4.683.066). RESULTS: The evaluation of SNOT-22 revealed that five patients showed an improvement in symptoms, while two remained in the same range of interpretation. One patient developed post-operative maxillary sinusitis, which was not detected at the time of evaluation by SNOT-22 or CBCT. CBCT showed a worsening sinus condition in three patients, two of whom had a significant increase in total bacteria count in their nasal cavities. The Brodsky scale was used to assess hypertrophy in palatine tonsils, where 60% of the subjects had grade 1 tonsils, 20% had grade 2 and 20% had grade 3. None of the patients had grade 4 tonsils, which would indicate more than 75% obstruction. Two patients harboured S. aureus and K. pneumoniae in their nasal cavities. Notably, K. pneumoniae, which was multidrug-resistant, was present in the nasal cavity of patients even before surgery, but this did not result in maxillary sinusitis, likely due to the patients' young and healthy condition. CONCLUSION: There was an improvement in signs and symptoms of maxillary sinusitis and quality of life in most patients after orthognathic surgery. However, some patients may still harbour multidrug-resistant bacteria, even if they are asymptomatic. Therefore, a thorough pre-operative assessment is essential to avoid difficult-to-treat post-operative complications.

Gut-associated microbes are present and active in the pig nasal cavity.

The nasal microbiota is a key contributor to animal health, and characterizing the nasal microbiota composition is an important step towards elucidating the role of its different members. Efforts to characterize the nasal microbiota composition of domestic pigs and other farm animals frequently report the presence of bacteria that are typically found in the gut, including many anaerobes from the Bacteroidales and Clostridiales orders. However, the in vivo role of these gut-microbiota associated taxa is currently unclear. Here, we tackled this issue by examining the prevalence, origin, and activity of these taxa in the nasal microbiota of piglets. First, analysis of the nasal microbiota of farm piglets sampled in this study, as well as various publicly available data sets, revealed that gut-microbiota associated taxa indeed constitute a substantial fraction of the pig nasal microbiota that is highly variable across individual animals. Second, comparison of herd-matched nasal and rectal samples at amplicon sequencing variant (ASV) level showed that these taxa are largely shared in the nasal and rectal microbiota, suggesting a common origin driven presumably by the transfer of fecal matter. Third, surgical sampling of the inner nasal tract showed that gut-microbiota associated taxa are found throughout the nasal cavity, indicating that these taxa do not stem from contaminations introduced during sampling with conventional nasal swabs. Finally, analysis of cDNA from the 16S rRNA gene in these nasal samples indicated that gut-microbiota associated taxa are indeed active in the pig nasal cavity. This study shows that gut-microbiota associated taxa are not only present, but also active, in the nasal cavity of domestic pigs, and paves the way for future efforts to elucidate the function of these taxa within the nasal microbiota.

Human nasal microbiota shifts in healthy and chronic respiratory disease conditions.

BACKGROUND: An increasing number of studies investigate various human microbiotas and their roles in the development of diseases, maintenance of health states, and balanced signaling towards the brain. Current data demonstrate that the nasal microbiota contains a unique and highly variable array of commensal bacteria and opportunistic pathogens. However, we need to understand how to harness current knowledge, enrich nasal microbiota with beneficial microorganisms, and prevent pathogenic developments. RESULTS: In this study, we have obtained nasal, nasopharyngeal, and bronchoalveolar lavage fluid samples from healthy volunteers and patients suffering from chronic respiratory tract diseases for full-length 16 S rRNA sequencing analysis using Oxford Nanopore Technologies. Demographic and clinical data were collected simultaneously. The microbiome analysis of 97 people from Lithuania suffering from chronic inflammatory respiratory tract disease and healthy volunteers revealed that the human nasal microbiome represents the microbiome of the upper airways well. CONCLUSIONS: The nasal microbiota of patients was enriched with opportunistic pathogens, which could be used as indicators of respiratory tract conditions. In addition, we observed that a healthy human nasal microbiome contained several plant- and bee-associated species, suggesting the possibility of enriching human nasal microbiota via such exposures when needed. These candidate probiotics should be investigated for their modulating effects on airway and lung epithelia, immunogenic properties, neurotransmitter content, and roles in maintaining respiratory health and nose-brain interrelationships.

Proteomic scrutiny of nasal microbiomes: implications for the clinic.

INTRODUCTION: The nasal cavity is the initial site of the human respiratory tract and is one of the habitats where microorganisms colonize. The findings from a growing number of studies have shown that the nasal microbiome is an important factor for human disease and health. 16S rRNA sequencing and metagenomic next-generation sequencing (mNGS) are the most commonly used means of microbiome evaluation. Among them, 16S rRNA sequencing is the primary method used in previous studies of nasal microbiomes. However, neither 16S rRNA sequencing nor mNGS can be used to analyze the genes specifically expressed by nasal microorganisms and their functions. This problem can be addressed by proteomic analysis of the nasal microbiome. AREAS COVERED: In this review, we summarize current advances in research on the nasal microbiome, introduce the methods for proteomic evaluation of the nasal microbiome, and focus on the important roles of proteomic evaluation of the nasal microbiome in the diagnosis and treatment of related diseases. EXPERT OPINION: The detection method for microbiome-expressed proteins is known as metaproteomics. Metaproteomic analysis can help us dig deeper into the nasal microbiomes and provide new targets and ideas for clinical diagnosis and treatment of many nasal dysbiosis-related diseases.

Prolonged mask wearing changed nasal microbial characterization of young adults during the COVID-19 pandemic in Shanghai, China.

Background: Face masks have become a common sight during the Coronavirus Disease 2019 (COVID-19) pandemic in many countries. However, the impact of prolonged face mask wearing on nasal microbiota of healthy people is not fully understood. Methods: In this study, we compared the nasal microbiota of 82 young adults who wore face masks for an extended period of time to 172 mask-free peers from the same school recruited before the COVID-19 pandemic via 16S ribosomal RNA gene sequencing. Diversity, composition, and function of nasal microbiota between the two groups were analyzed. Prevalence of commensal bacteria colonized in the nasal cavity was determined by culture-based analysis. Results: We observed that prolonged face mask wearers had significantly different nasal microbial characterization and metabolic function compared to mask-free controls from 2018. Specifically, the nasal microbiota of the prolonged mask wearers displayed increased abundance of Staphylococcus, Pseudoalteromonas, Corynebacterium, etc. Meanwhile, the abundance of several genera including Bacteroides, Faecalibacterium, and Agathobacter was decreased. Moreover, we observed that COVID-19 infection history did not affect the composition of nasal microbiota significantly. Additionally, the culture-based analysis revealed that Staphylococcus aureus and Corynebacterium accolens increased, and Staphylococcus epidermidis decreased in the nasal cavity of prolonged mask wearers. Conclusions: Overall, our study suggests that prolonged face mask wearing can significantly alter the nasal microbiota.

Limited Adaptation of Staphylococcus aureus during Transition from Colonization to Invasive Infection.

Staphylococcus aureus carriage is a risk factor for invasive infections. Unique genetic elements favoring the transition from colonizing to invasive phenotype have not yet been identified, and phenotypic adaptation traits are understudied. We therefore assessed phenotypic and genotypic profiles of 11 S. aureus isolate pairs sampled from colonized patients simultaneously suffering from invasive S. aureus infections. Ten out of 11 isolate pairs displayed the same spa and multilocus sequence type, suggesting colonization as an origin for the invasive infection. Systematic analysis of colonizing and invasive isolate pairs showed similar adherence, hemolysis, reproductive fitness properties, antibiotic tolerance, and virulence in a Galleria mellonella infection model, as well as minimal genetic differences. Our results provide insights into the similar phenotypes associated with limited adaptation between colonizing and invasive isolates. Disruption of the physical barriers of mucosa or skin was identified in the majority of patients, further emphasizing colonization as a major risk factor for invasive disease. IMPORTANCE S. aureus is a major pathogen of humans, causing a wide range of diseases. The difficulty to develop a vaccine and antibiotic treatment failure warrant the exploration of novel treatment strategies. Asymptomatic colonization of the human nasal passages is a major risk factor for invasive disease, and decolonization procedures have been effective in preventing invasive infections. However, the transition of S. aureus from a benign colonizer of the nasal passages to a major pathogen is not well understood, and both host and bacterial properties have been discussed as being relevant for this behavioral change. We conducted a thorough investigation of patient-derived strain pairs reflecting colonizing and invasive isolates in a given patient. Although we identified limited genetic adaptation in certain strains, as well as slight differences in adherence capacity among colonizing and invasive isolates, our work suggests that barrier breaches are a key event in the disease continuum of S. aureus.

Microbial characterization of the nasal cavity in patients with allergic rhinitis and non-allergic rhinitis.

Introduction: Although recent studies have shown that the human microbiome is involved in the pathogenesis of allergic diseases, the impact of microbiota on allergic rhinitis (AR) and non-allergic rhinitis (nAR) has not been elucidated. The aim of this study was to investigate the differences in the composition of the nasal flora in patients with AR and nAR and their role in the pathogenesis. Method: From February to September 2022, 35 AR patients and 35 nAR patients admitted to Harbin Medical University's Second Affiliated Hospital, as well as 20 healthy subjects who underwent physical examination during the same period, were subjected to 16SrDNA and metagenomic sequencing of nasal flora. Results: The microbiota composition of the three groups of study subjects differs significantly. The relative abundance of Vibrio vulnificus and Acinetobacter baumanni in the nasal cavity of AR patients was significantly higher when compared to nAR patients, while the relative abundance of Lactobacillus murinus, Lactobacillus iners, Proteobacteria, Pseudomonadales, and Escherichia coli was lower. In addition, Lactobacillus murinus and Lacttobacillus kunkeei were also negatively correlated with IgE, while Lacttobacillus kunkeei was positively correlated with age. The relative distribution of Faecalibacterium was higher in moderate than in severe AR patients. According to KEGG functional enrichment annotation, ICMT(protein-S-isoprenylcysteine O-methyltransferase,ICMT) is an AR microbiota-specific enzyme that plays a role, while glycan biosynthesis and metabolism are more active in AR microbiota. For AR, the model containing Parabacteroides goldstemii, Sutterella-SP-6FBBBBH3, Pseudoalteromonas luteoviolacea, Lachnospiraceae bacterium-615, and Bacteroides coprocola had the highest the area under the curve (AUC), which was 0.9733(95%CI:0.926-1.000) in the constructed random forest prediction model. The largest AUC for nAR is 0.984(95%CI:0.949-1.000) for the model containing Pseudomonas-SP-LTJR-52, Lachnospiraceae bacterium-615, Prevotella corporis, Anaerococcus vaginalis, and Roseburia inulinivorans. Conclusion: In conclusion, patients with AR and nAR had significantly different microbiota profiles compared to healthy controls. The results suggest that the nasal microbiota may play a key role in the pathogenesis and symptoms of AR and nAR, providing us with new ideas for the treatment of AR and nAR.

Staphylococcus aureus nasal colonization level and intracellular reservoir: a prospective cohort study.

Staphylococcus aureus is a major pathogen in humans. The nasal vestibule is considered as the main reservoir of S. aureus. However, even though the nasal cavity may also be colonized by S. aureus, the relationships between the two sites are still unclear. We conducted a prospective study in humans to assess the S. aureus colonization profiles in the vestibule and nasal cavity, and to investigate the presence of intracellular S. aureus in the two sites. Patients undergoing ear, nose, and throat surgery were swabbed during endoscopy to determine S. aureus nasal load, genotype, and presence of intracellular S. aureus. Among per-operative samples from 90 patients, the prevalence of S. aureus carriage was 32.2% and 33.3% in the vestibule and the nasal cavity, respectively. The mean S. aureus load was 4.10 and 4.25 log10 CFU/swab for the nasal vestibule and nasal cavity, respectively (P > 0.05). Genotyping of S. aureus revealed that all nasal strains isolated from a given individual belong to the same clonal complex and spa-type. An intracellular carriage was observed in 5.6% of the patients, all of whom exhibited a S. aureus vestibule load higher than 3 log10 CFU/swab. An intracellular niche was observed in the vestibule as well as in the nasal cavity. In conclusion, the nasal cavity was also found to be a major site of S. aureus carriage in humans and should draw attention when studying host-pathogen interactions related to the risk of infection associated with colonization.

Prevalencia de portación nasal de Staphylococcus aureus sensible y resistente a la meticilina en candidatos a artroplastia total de cadera o rodilla / Prevalence of nasal carriage of methicillin-sensitive and methicillin-resistant Staphylococcus aureus in candidates for total hip or knee arthroplasty

INTRODUCIÓN Las infecciones perioperatorias en cirugía de reemplazo articular son fuente importante de morbimortalidad, así como de altos costos económicos y sociales, tanto para el paciente como para su entorno. La colonización preoperatoria por Staphylococcus aureus ha sido reconocida como un factor de riesgo importante para desarrollar una infección de sitio quirúrgico.El objetivo de este estudio es conocer la prevalencia de portación nasal de S. aureus, tanto sensible a la meticilina (SASM) como resistente a la meticilina (SARM), en pacientes candidatos a cirugía de reemplazo articular de cadera o rodilla. MATERIALES Y MÉTODOS Se realizó un estudio observacional de una cohorte retrospectiva de pacientes con indicación de artroplastia total de cadera (ATC) y rodilla (ATR) electiva por artrosis severa en un hospital público de Chile. Los pacientes fueron sometidos a tamizaje preoperatorio de portación, cultivándose muestras obtenidas mediante hisopado de ambas fosas nasales. Los datos del laboratorio fueron recopilados y presentados como porcentaje de portación de S. aureus. RESULTADOS Se estudiaron 303 pacientes consecutivos de ATC y 343 de ATR. En total, 483 de los 646 pacientes (74,7%) tuvieron estudio preoperatorio de portación nasal. Se identificaron 123 pacientes (25,4%) portadores de S. aureus, de los cuales sólo 2 (0,41%) casos correspondieron a SARM. CONCLUSIÓN La prevalencia de portación nasal de S. aureus obtenida fue de 25%, similar a lo reportado en otras series. La prevalencia de SARM (0.41%), sin embargo, estuvo bajo lo descrito en la literatura internacional (0,6­6%). Sería de utilidad, dada la alta prevalencia de portación descrita en nuestro trabajo y de acuerdo a evidencia publicada recientemente, realizar protocolos de descolonización universales, sin necesidad de realizar tamizaje preoperatorio.

INTRODUCTION Surgical-site infections in joint replacement surgery are an important source of morbidity and mortality that entail high economic and social burden both for the patient and their environment. Preoperative colonization by Staphylococcus aureus has been recognized as an important risk factor for the development of surgical-site infection. The aim of the present study is to determine the prevalence of nasal colonization by S. aureus, both methicillin-sensitive (MSSA) and methicillin-resistant (MRSA) in patients who are candidates for total replacement of the hip or knee joints. MATERIALS AND METHODS A retrospective observational study of a cohort of 646 patients with an indication to undergo total hip arthroplasty (THA) or total knee arthroplasty (TKA) due to severe osteoarthritis was performed in a Public Hospital in Chile. The patients were submitted to a preoperative screening for S. aureus carriage, and the culture samples were obtained by swabbing both nostrils. The laboratory data was collected and presented as a percentage of carriage. RESULTS We consecutively examined 303 THA and 343 TKA patients. A total of 483 of the 646 patients (74.7%) underwent a preoperative study of nasal carriage. We identified 123 (25.4%) S. aureus carriers, and only found 2 (0.41%) cases corresponding to MRSA. CONCLUSION We found a prevalence of nasal carriage of S. aureus of 25.4%, a rate similar to that reported in other series. The prevalence of MRSA (0.41%), however, was lower than that reported in the international literature (0.6­6%). Given the high prevalence of carriage described in our work and according to recently published data, it would be worthwhile to carry out universal decolonization protocols, without the need for preoperative screening.

Nasal microbiota evolution within the congregate setting imposed by military training.

The human microbiome is comprised of a complex and diverse community of organisms that is subject to dynamic changes over time. As such, cross-sectional studies of the microbiome provide a multitude of information for a specific body site at a particular time, but they fail to account for temporal changes in microbial constituents resulting from various factors. To address this shortcoming, longitudinal research studies of the human microbiome investigate the influence of various factors on the microbiome of individuals within a group or community setting. These studies are vital to address the effects of host and/or environmental factors on microbiome composition as well as the potential contribution of microbiome members during the course of an infection. The relationship between microbial constituents and disease development has been previously explored for skin and soft tissue infections (SSTIs) within congregate military trainees. Accordingly, approximately 25% of the population carries Staphylococcus aureus within their nasal cavity, and these colonized individuals are known to be at increased risk for SSTIs. To examine the evolution of the nasal microbiota of U.S. Army Infantry trainees, individuals were sampled longitudinally from their arrival at Fort Benning, Georgia, until completion of their training 90 days later. These samples were then processed to determine S. aureus colonization status and to profile the nasal microbiota using 16S rRNA gene-based methods. Microbiota stability differed dramatically among the individual trainees; some subjects exhibited great stability, some subjects showed gradual temporal changes and some subjects displayed a dramatic shift in nasal microbiota composition. Further analysis utilizing the available trainee metadata suggests that the major drivers of nasal microbiota stability may be S. aureus colonization status and geographic origin of the trainees. Nasal microbiota evolution within the congregate setting imposed by military training is a complex process that appears to be affected by numerous factors. This finding may indicate that future campaigns to prevent S. aureus colonization and future SSTIs among high-risk military trainees may require a 'personalized' approach.

Portación nasal de Staphylococcus aureus en el personal de salud de áreas críticas de un Hospital Pediátrico durante julio-setiembre 2018 / Nasal carriage of Staphylococcus aureus in health personnel in critical areas of a Pediatric Hospital during July-September 2018 / Transporte nasal de Staphylococcus aureus em profissionais de saúde em áreas críticas de um Hospital Pediátrico durante julho-setembro de 2018

Las infecciones intrahospitalarias (IIH) son causa de elevada morbimortalidad y representan un problema sanitario importante. El personal de salud es reservorio y potencial transmisor de los agentes etiológicos de las mismas. S. aureus es uno de los microorganismos implicados, por lo tanto es importante conocer la frecuencia de portación en el personal de salud y establecer el perfil de susceptibilidad antimicrobiana para contribuir con la elaboración de medidas de prevención incluyendo actividades educativas. Objetivo: Conocer la frecuencia de portación de S. aureus, distribución y antibiotipos de las cepas presentes en el personal sanitario del Hospital Pediátrico de Referencia (HPR). Materiales y métodos: Se realizó un estudio descriptivo durante el periodo julio-setiembre del año 2018. Se incluyeron muestras de hisopados nasales de trabajadores de la salud de distintas áreas de internación que consintieron participar en el estudio. Se excluyeron aquellos que recibieron antibióticos dentro de los 3 meses previos al estudio. Las muestras fueron sembradas en agar sangre ovina al 5% (ASO) y se incubaron a 35-37ºC en aerobiosis por 24-48 horas. La identificación de las colonias sospechosas de Staphylococcus aureus por métodos convencionales y MALDI-TOF. El patrón de resistencia antimicrobiana de S. aureus se detectó por disco-difusión. En los cultivos resistentes a meticilina (SAMR) se determinó la presencia del gen mecA y se realizó la tipificación del SCCmec por pruebas de reacción en cadena de polimerasa. Resultados: Se obtuvieron 225 hisopados a partir de 225 trabajadores, presentaron desarrollo 212. En 49 se recuperaron cultivos de S. aureus. Correspondieron a SAMR 11 de las 49 cepas, todas portaban el gen mecA. Hubo predominio en el personal de enfermería (7/11), en los servicios de hemato-oncología (3/11) y cuidados intensivos neonatales (4/11). Asociaron resistencia a macrólidos y clindamicina 8 de 11 aislamientos SAMR, a gentamicina 2 y a mupirocina uno. El SCCmec más frecuentemente identificado fue el tipo IV (7/11). Conclusiones: Los resultados muestran la presencia de cepas SAMR entre el personal de salud del CHPR y aportan información complementaria para efectuar prevención y control de las IIH, actuando sobre todo en el personal de salud encargado de la atención de pacientes susceptibles.

Hospital-acquired infections (IIH) are a cause of high morbidity and mortality and represent a major health problem. Health personnel are reservoirs and potential transmitters of their etiological agents. S. aureus is one of the microorganisms involved, therefore it is important to know the frequency of carriage in health personnel and establish the antimicrobial susceptibility profile to contribute to the development of prevention measures, including educational activities. Objective: To know the frequency of carriage of S. aureus, distribution and antibiotypes of the strains present in the health personnel of the Reference Pediatric Hospital (HPR). Materials and methods: A descriptive study was carried out during the period July-September 2018. Nasal swab samples from health workers from different hospitalization areas who agreed to participate in the study were included. Those who received antibiotics within 3 months prior to the study were excluded. The samples were seeded in 5% sheep blood agar (ASO) and incubated at 35-37ºC in aerobiosis for 24-48 hours. Identification of suspicious Staphylococcus aureus colonies by conventional methods and MALDI-TOF. The antimicrobial resistance pattern of S. aureus was detected by disc diffusion. In methicillin-resistant cultures (MRSA), the presence of the mecA gene was determined and SCCmec was typified by polymerase chain reaction tests. Results: 225 swabs were obtained from 225 workers, 212 showed development. S. aureus cultures were recovered from 49. 11 of the 49 strains corresponded to MRSA, all of them carried the mecA gene. There was a predominance in the nursing staff (7/11), in the hematology-oncology services (3/11) and neonatal intensive care (4/11). They associated resistance to macrolides and clindamycin in 8 of 11 MRSA isolates, 2 to gentamicin, and 1 to mupirocin. The most frequently identified SCCmec was type IV (7/11). Conclusions: The results show the presence of MRSA strains among the health personnel of the CHPR and provide complementary information to carry out prevention and control of IIH, acting especially on the health personnel in charge of the care of susceptible patients.

As infecções hospitalares (HII) são causa de alta morbidade e mortalidade e representam um importante problema de saúde. Os profissionais de saúde são reservatórios e potenciais transmissores de seus agentes etiológicos. O S. aureus é um dos micro-organismos envolvidos, por isso é importante conhecer a frequência de portadores em profissionais de saúde e estabelecer o perfil de suscetibilidade antimicrobiana para contribuir no desenvolvimento de medidas de prevenção incluindo atividades educativas. Objetivo: Conhecer a frequência de portadores de S. aureus, distribuição e antibiótipos das cepas presentes no pessoal de saúde do Hospital Pediátrico de Referência (HPR). Materiais e métodos: Foi realizado um estudo descritivo durante o período de julho a setembro de 2018. Foram incluídas amostras de swab nasal de profissionais de saúde de diferentes áreas de internação que concordaram em participar do estudo. Aqueles que receberam antibióticos nos 3 meses anteriores ao estudo foram excluídos. As amostras foram semeadas em 5% de ágar sangue de carneiro (ASO) e incubadas a 35-37ºC em aerobiose por 24-48 horas. Identificação de colônias suspeitas de Staphylococcus aureus por métodos convencionais e MALDI-TOF. O padrão de resistência antimicrobiana de S. aureus foi detectado por difusão em disco. Em culturas resistentes à meticilina (MRSA), a presença do gene mecA foi determinada e SCCmec foi tipificado por testes de reação em cadeia da polimerase. Resultados: 225 swabs foram obtidos de 225 trabalhadores, 212 apresentaram desenvolvimento. Culturas de S. aureus foram recuperadas de 49. 11 das 49 cepas correspondiam a MRSA, todas carregavam o gene mecA. Houve predominância na equipe de enfermagem (7/11), nos serviços de hematologia-oncologia (3/11) e de terapia intensiva neonatal (4/11). Eles associaram resistência a macrolídeos e clindamicina em 8 de 11 isolados de MRSA, 2 à gentamicina e 1 à mupirocina. O SCCmec mais frequentemente identificado foi o tipo IV (7/11). Conclusões: Os resultados mostram a presença de cepas de MRSA entre os profissionais de saúde do CHPR e fornecem informações complementares para realizar a prevenção e controle da HII, atuando principalmente sobre os profissionais de saúde responsáveis ​​pelo atendimento de pacientes suscetíveis.

The Fim and FhaB adhesins play a crucial role in nasal cavity infection and Bordetella pertussis transmission in a novel mouse catarrhal infection model.

Pulmonary infections caused by Bordetella pertussis used to be the prime cause of infant mortality in the pre-vaccine era and mouse models of pertussis pneumonia served in characterization of B. pertussis virulence mechanisms. However, the biologically most relevant catarrhal disease stage and B. pertussis transmission has not been adequately reproduced in adult mice due to limited proliferation of the human-adapted pathogen on murine nasopharyngeal mucosa. We used immunodeficient C57BL/6J MyD88 KO mice to achieve B. pertussis proliferation to human-like high counts of 108 viable bacteria per nasal cavity to elicit rhinosinusitis accompanied by robust shedding and transmission of B. pertussis bacteria to adult co-housed MyD88 KO mice. Experiments with a comprehensive set of B. pertussis mutants revealed that pertussis toxin, adenylate cyclase toxin-hemolysin, the T3SS effector BteA/BopC and several other known virulence factors were dispensable for nasal cavity infection and B. pertussis transmission in the immunocompromised MyD88 KO mice. In contrast, mutants lacking the filamentous hemagglutinin (FhaB) or fimbriae (Fim) adhesins infected the nasal cavity poorly, shed at low levels and failed to productively infect co-housed MyD88 KO or C57BL/6J mice. FhaB and fimbriae thus appear to play a critical role in B. pertussis transmission. The here-described novel murine model of B. pertussis-induced nasal catarrh opens the way to genetic dissection of host mechanisms involved in B. pertussis shedding and to validation of key bacterial transmission factors that ought to be targeted by future pertussis vaccines.

Colonization of nasal cavities by Staphylococcus epidermidis mitigates SARS-CoV-2 nucleocapsid phosphoprotein-induced interleukin (IL)-6 in the lung.

Infection by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can trigger excessive interleukin (IL)-6 signalling, leading to a myriad of biological effects including a cytokine storm that contributes to multiple organ failure in severe coronavirus disease 2019 (COVID-19). Using a mouse model, we demonstrated that nasal inoculation of nucleocapsid phosphoprotein (NPP) of SARS-CoV-2 increased IL-6 content in bronchoalveolar lavage fluid (BALF). Nasal administration of liquid coco-caprylate/caprate (LCC) onto Staphylococcus epidermidis (S. epidermidis)-colonized mice significantly attenuated NPP-induced IL-6. Furthermore, S. epidermidis-mediated LCC fermentation to generate electricity and butyric acid that promoted bacterial colonization and activated free fatty acid receptor 2 (Ffar2) respectively. Inhibition of Ffar2 impeded the effect of S. epidermidis plus LCC on the reduction of NPP-induced IL-6. Collectively, these results suggest that nasal S. epidermidis is part of the first line of defence in ameliorating a cytokine storm induced by airway infection of SARS-CoV-2.

Macrococcus armenti sp. nov., a novel bacterium isolated from the skin and nasal cavities of healthy pigs and calves.

Gram-positive coccoid bacteria were isolated from the nasal cavities of pigs and calves as well as from axillar and inguinal skin regions of pigs. Phylogenetic analysis of seven strains based on complete genome, 16S rRNA, hsp60, dnaJ, rpoB and sodA gene sequences and MALDI-TOF MS profiles revealed that they belonged to the genus Macrococcus with the closest relatedness to Macrococcus canis, Macrococcus caseolyticus subsp. caseolyticus and Macrococcus caseolyticus subsp. hominis. DNA relatedness of the type strain JEK37T with the type strains of M. canis, M. caseolyticus subsp. caseolyticus and M. caseolyticus subsp. hominis was 23.4, 23.1 and 23.0 % by digital DNA-DNA hybridization and 80.39, 80.45 and 80.87 % by average nucleotide identity (ANI) calculations, confirming that they do not belong to the same species. The DNA G+C content of JEK37T was 35.65 mol%. The novel strains can be differentiated from M. canis KM 45013T by the ability to fermentate d-ribose and by the absence of DNAase production and haemolysis, from M. caseolyticus subsp. caseolyticus CCUG 15606T by the ability to fermentate sucrose and from both species by the inability to grow in 9 and 12% NaCl. They differ from M. caseolyiticus subsp. hominis by the presence of α-glucosidase. The most common fatty acids of JEK37T were C14 : 0, C18 : 1 ω9c and C18 : 0. Known polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, aminolipid, aminoglycolipid, aminophospholipid, glycolipid and phospholipid. Cell-wall peptidoglycan of JEK37T was of type A3α l-Lys-Gly2-L-Ser-Gly (similar to A11.3) and the respiratory quinolone was menaquinone 6. Based on their genotypic and chemotaxonomic characteristics, these strains represent a novel species of the genus Macrococcus, for which we propose the name Macrococcus armenti sp. nov. The type strain is JEK37T (=DSM 112712T=CCOS 1982T).

Detection of Mycobacterium tuberculosis complex DNA in oronasal swabs from infected African buffaloes (Syncerus caffer).

Mycobacterium bovis (M. bovis), a member of the Mycobacterium tuberculosis complex (MTBC), is the causative agent of bovine TB (bTB) in animals. Spread occurs through inhalation or ingestion of bacilli transmitted from infected individuals. Early and accurate detection of infected African buffaloes shedding M. bovis is essential for interrupting transmission. In this pilot study, we determined if MTBC DNA could be detected in M. bovis infected buffalo oronasal secretions using a molecular transport media (PrimeStore MTM) with oronasal swabs and a rapid qPCR assay (Xpert MTB/RIF Ultra). Bovine TB test-positive buffaloes were culled, then tissue samples and oronasal swabs collected post-mortem for mycobacterial culture and Ultra testing, respectively. The Ultra detected MTBC DNA in 5/12 swabs from M. bovis culture-confirmed buffaloes. Oronasal swabs from M. bovis negative buffaloes (n = 20) were negative on Ultra, indicating the high specificity of this test. This study showed that MTM can successfully preserve MTBC DNA in oronasal swabs. The proportion of MTBC positive oronasal swabs was higher than expected and suggests that the Ultra may be an additional method for identifying infected buffaloes. Further studies are needed to confirm the utility of the Ultra assay with oronasal swabs as an assay to evaluate possible MTBC shedding in buffaloes.

Identification and Characterization of a Two-Peptide Class IIb Bacteriocin in Streptococcus pluranimalium Isolated from the Nasal Cavity of a Healthy Pig.

In addition to be an important zoonotic agent, Streptococcus suis serotype 2 causes severe infections in pigs. In this study, we characterized a new bacteriocin produced by Streptococcus pluranimalium 2N12 isolated from a pig nasal sample. The bacteriocin, termed pluranimalicin 2N12, was a two-peptide class IIb bacteriocin active against S. suis. The gene cluster responsible for the biosynthesis of pluranimalicin 2N12 by S. pluranimalium contained seven open reading frames, including putative genes for peptides (pluα, pluß), export (pluA, pluB), and regulation (pluC, pluD, pluE). The deduced amino acid sequences of the peptides Pluα (33 amino acids) and Pluß (29 amino acids) showed 73% and 69% identity in amino acid residues, respectively, with the peptides SthA and SthB of the streptocin produced by Streptococcus gordonii. The antibacterial activity of pluranimalicin 2N12 against S. suis was dependent on the presence of the two peptides Pluα and Pluß that exhibited a membrane permeabilization effect. No activity was found against the other swine pathogens tested. Depending on the concentrations used, Pluα and Pluß displayed no or low toxicity towards swine tracheal epithelial cells. The pluranimalicin peptides Pluα and Pluß, either individually or in combination, exhibited anti-inflammatory activity since they attenuated IL-6 and TNF-α production by macrophages challenged with lipopolysaccharide. Given its dual action (antibacterial and anti-inflammatory), pluranimalicin 2N12 holds promise as a potential therapeutic agent for controlling S. suis infections.

A Lateral Flow Test for Staphylococcus aureus in Nasal Mucus Using a New DNAzyme as the Recognition Element.

Detection of pathogenic bacteria in complex biological matrices remains a major challenge. Herein, we report the selection and optimization of a new DNAzyme for Staphylococcus aureus (SA) and the use of the DNAzyme to develop a simple lateral flow device (LFD) for detection of SA in nasal mucus. The DNAzyme was generated by in vitro selection using a crude extra/intracellular mixture derived from SA, which could be used directly for simple solution or paper-based fluorescence assays for SA. The DNAzyme was further modified to produce a DNA cleavage fragment that acted as a bridging element to bind DNA-modified gold nanoparticles to the test line of a LFD, producing a simple colorimetric dipstick test. The LFD was evaluated with nasal mucus samples spiked with SA, and demonstrated that SA detection was possible in minutes with minimal sample processing.

Methicillin-resistant Staphylococcus aureus colonization among healthcare workers in Oman.

INTRODUCTION: Methicillin-Resistant Staphylococcus aureus (MRSA) is a S. aureus strain characterized by resistance to cloxacillin. Healthcare workers (HCWs), are recognized for their heightened risk for MRSA acquisition and possibly for MRSA nosocomial transmission. This cross-sectional study aimed to determine the prevalence and the associated risk factors of MRSA colonization among healthcare workers at Sultan Qaboos University Hospital (SQUH) in Oman. METHODOLOGY: A total of 200 nasal swab samples were collected from the healthcare workers at SQUH during the period October 2nd 2018 to January 7th 2019. All nasal swab samples were examined microbiologically for the presence of MRSA using the standard method and the results were confirmed by detection of the mecA product (PBP2a). Data on associated risk factors for MRSA colonization was collected and analyzed. RESULTS: Forty-one of the 200 screened healthcare workers (20.5%) were found to have nasal carriage of Staphylococcus aureus of which 63.4% were Methicillin Sensitive and 36.6% were Methicillin-Resistant (MRSA). Methicillin-Resistant Staphylococcus aureus (MRSA) was isolated from fifteen of the 200 screened healthcare workers giving a prevalence rate of nasal colonization with MRSA of 7.5%. We found no statistical association between healthcare worker MRSA nasal colonization and age, gender, HCWs specialty, hand hygiene practices, skin condition, previous MRSA infection, and previous exposure to antibiotics. CONCLUSIONS: Identification of the prevalence and the associated risk factors of MRSA colonization in healthcare workers mandates continuous surveillance and the implementation of all possible preventive measures to reduce re-occurrences.

Upper Respiratory Tract Microbiome of Australian Aboriginal and Torres Strait Islander Children in Ear and Nose Health and Disease.

The objective of this study was to examine the nasal microbiota in relation to otitis media (OM) status and nose health in Indigenous Australian children. Children 2 to 7 years of age were recruited from two northern Australian (Queensland) communities. Clinical histories were obtained through parent interviews and reviews of the medical records. Nasal cavity swab samples were obtained, and the children's ears, nose, and throat were examined. DNA was extracted and analyzed by 16S rRNA amplicon next-generation sequencing of the V3/V4 region, in combination with previously generated culture data. A total of 103 children were recruited (mean age, 4.7 years); 17 (16.8%) were healthy, i.e., normal examination results and no history of OM. The nasal microbiota differed significantly in relation to OM status and nose health. Children with historical OM had greater relative abundance of Moraxella, compared to healthy children, despite both having healthy ears at the time of swabbing. Children with healthy noses had greater relative abundance of Staphylococcus aureus, compared to those with rhinorrhea. Dolosigranulum was correlated with Corynebacterium in healthy children. Haemophilus and Streptococcus were correlated across phenotypes. Ornithobacterium was absent or was present with low relative abundance in healthy children and clustered around otopathogens. It correlated with Helcococcus and Dichelobacter. Dolosigranulum and Corynebacterium form a synergism that promotes upper respiratory tract (URT)/ear health in Indigenous Australian children. Ornithobacterium likely represents "Candidatus Ornithobacterium hominis" and in this population is correlated with a novel bacterium that appears to be related to poor URT/ear health. IMPORTANCE Recurring and chronic infections of the ear (OM) are disproportionately prevalent in disadvantaged communities across the globe and, in particular, within Indigenous communities. Despite numerous intervention strategies, OM persists as a major health issue and is the leading cause of preventable hearing loss. In disadvantaged communities, this hearing loss is associated with negative educational and social development outcomes, and consequently, poorer employment prospects and increased contact with the justice system in adulthood. Thus, a better understanding of the microbial ecology is needed in order to identify new targets to treat, as well as to prevent the infections. This study used a powerful combination of 16S rRNA gene sequencing and extended culturomics to show that Dolosigranulum pigrum, a bacterium previously identified as a candidate protective species, may require cocolonization with Corynebacterium pseudodiphtheriticum in order to prevent OM. Additionally, emerging and potentially novel pathogens and bacteria were identified.