Genome-Scale Characterization of Mycobacterium abscessus Complex Isolates from Portugal.

The Mycobacterium abscessus complex (MABC) is an emerging, difficult to treat, multidrug-resistant nontuberculous mycobacteria responsible for a wide spectrum of infections and associated with an increasing number of cases worldwide. Dominant circulating clones (DCCs) of MABC have been genetically identified as groups of strains associated with higher prevalence, higher levels of antimicrobial resistance, and worse clinical outcomes. To date, little is known about the genomic characteristics of MABC species circulating in Portugal. Here, we examined the genetic diversity and antimicrobial resistance profiles of 30 MABC strains isolated between 2014 and 2022 in Portugal. The genetic diversity of circulating MABC strains was assessed through a gene-by-gene approach (wgMLST), allowing their subspecies differentiation and the classification of isolates into DCCs. Antimicrobial resistance profiles were defined using phenotypic, molecular, and genomic approaches. The majority of isolates were resistant to at least two antimicrobials, although a poor correlation between phenotype and genotype data was observed. Portuguese genomes were highly diverse, and data suggest the existence of MABC lineages with potential international circulation or cross-border transmission. This study highlights the genetic diversity and antimicrobial resistance profile of circulating MABC isolates in Portugal while representing the first step towards the implementation of a genomic-based surveillance system for MABC at the Portuguese NIH.

Microevolution of a Mycobacteroides abscessus subsp. bolletii strain in a clinical persistent infection.

Mycobacteroides abscessus complex (MAB), a fast-growing nontuberculous mycobacterium, is emerging as a significant infectious disease threat, due to both intrinsic and acquired resistance mechanisms to antibiotics and disinfectants and the need for extensive and multidrug regimens for treatment. Despite the prolonged regimens, outcomes are poor and persistence cases have been reported. Here, we describe clinical, microbiologic and genomic features of a M. abscessus subsp. bolletii (M. bolletii) strain consecutively isolated from a patient within an eight-year infection period. From April 2014 to September 2021, the National Reference Laboratory for Mycobacteria received eight strains isolated from a male patient. Species identification, molecular resistance profile and phenotypic drug susceptibility were determined. Five of these isolates were recovered for further in-depth genomic analysis. Genomic analysis confirmed the multidrug resistant pattern of the strain and also other genetic changes associated with adaptation to environment and defence mechanisms. We highlight the identification of new mutations in locus MAB_1881c and in locus MAB_4099c (mps1 gene), already described as associated with macrolides resistance and morphotype switching, respectively. Additionally, we also observed the emergence and fixation of a mutation in locus MAB_0364c that appeared at a frequency of 36% for the 2014 isolate, 57% for the 2015 isolate and 100% for the 2017 and 2021 isolates, clearly illustrating a fixation process underlying a microevolution of the MAB strain within the patient. Altogether these results suggest that the observed genetic alterations are a reflection of the bacterial population's continuous adaptation and survival to the host environment during infection, contributing to persistence and treatment failure.

Anonymized Distributed PHR Using Blockchain for Openness and Non-Repudiation Guarantee.

We introduce our solution developed for data privacy, and specifically for cognitive security that can be enforced and guaranteed using blockchain technology in SAAL (Smart Ambient Assisted Living) environments. Personal clinical and demographic information segments to various levels that assures that it can only be rebuilt at the interested and authorized parties and no profiling can be extracted from the blockchain itself. Using our proposal the access to a patient's clinical process resists tampering and ransomware attacks that have recently plagued the HIS (Hospital Information Systems) in various countries. The core of the blockchain model assures non-repudiation possible by any of the involved information producers thus maintaining ledger fidelity of the enclosed historical process information. One important side effect of this data infrastructure is that it can be accessed in open form, for research purposes for instance, since no individual re-identification or group profiling is possible by any means.

Análise da eficácia antimicrobiana do ácido peracético na desinfecção de moldes de hidrocoloide irreversível / Analysis of the antimicrobial effectiveness of peracetic acid for the disinfection of irreversible hydrocolloid impressions

Introdução: Nos últimos anos, o ácido peracético tem sido utilizado na Odontologia para desinfecção de moldes de hidrocoloide irreversível. Objetivo: Avaliar a eficácia antimicrobiana do ácido peracético nas técnicas de nebulização e pulverização, em moldes de hidrocoloide irreversível. Material e método: Oitenta moldes de hemiarcos de voluntários foram distribuídos aleatoriamente em quatro grupos experimentais (n=20), com respectivos controles. Os moldes foram submetidos à desinfecção por dez minutos com ácido peracético 0,2% e hipoclorito de sódio 1%, pelos métodos de nebulização, com atmosfera de 100% de umidade relativa, e pulverização. Após desinfecção, cada molde foi imerso em solução salina e submetido à vibração ultrassônica para dispersão de possíveis microrganismos. Para análise microbiológica desta solução, foi realizada contagem de colônias que cresceram em meio de cultura estéril BHI-ágar após 24 h em estufa incubadora a 37°C. Resultado: Em todos os grupos, foi significante a diferença no número médio de colônias decorrentes da análise estatística do controle e do grupo experimental correspondente (p<0,05). Comparando-se o ácido peracético (0,2%) nos dois métodos, não houve diferença significativa (p=0,420); no entanto, comparando-se o hipoclorito de sódio (1%) nos dois métodos, a diferença foi estatisticamente significante (p=0,010). No mesmo método e com soluções diferentes, a nebulização com ácido peracético 0,2% foi mais eficaz que o hipoclorito de sódio 1% (p=0,030). Conclusão: Ácido peracético 0,2% pode ser usado para desinfecção de moldes de hidrocoloide irreversível nos métodos de pulverização e nebulização. Hipoclorito de sódio 1% mostrou ser mais eficaz quando se usou o método de pulverização.

Introduction: In recent years, peracetic acid has been used in dentistry for disinfection of irreversible hydrocolloid. Objective: To evaluate the antimicrobial effectiveness of peracetic acid using nebulization and spray techniques on irreversible hydrocolloid impressions. Material and method: 80 impressions of the hemi-arches of volunteers were randomly divided into four experimental groups (n = 20), with respective controls. The impressions were disinfected for 10 minutes with 0.2% peracetic acid and 1% sodium hypochlorite using the methods of nebulization in a 100% relative humidity atmosphere, and spraying. After disinfection, each impression was immersed in saline solution and subjected to ultrasonic vibration to disperse possible microorganisms. For the microbiological analysis of the solution, bacterial colonies, grown in sterile BHI-agar culture after 24h in an oven incubator at 37 ° C, were counted. Result: There was a statistically significant difference between the mean number of colonies of the control and each experimental group (p <0.05). There was no statistically significant difference between the two methods when using 0.2% peracetic acid (p = 0.420). However, there was a statistically significant difference between the two methods when using 1% sodium hypochlorite (p = 0.010). Nebulization with 0.2% peracetic acid was more effective than 1% sodium hypochlorite (p=0.030). Conclusion: 0.2% peracetic acid can be used for disinfection of irreversible hydrocolloid impressions using the methods of spraying or nebulization. Sodium hypochlorite (1%) proved to be more effective using the spraying method.

Non-tuberculous mycobacteria in HIV-negative patients with pulmonary disease in Lisbon, Portugal.

Non-tuberculous mycobacteria (NTM) are responsible for an increase in mycobacterial disease in many developed countries. However the epidemiology, especially in patients without HIV infection, remains difficult to determine. We studied a convenience sample of 5497 HIV-negative patients receiving care at our pneumology centres. Among 510 patients with mycobacterial isolates, NTM were demonstrated in 58, all showing signs of lung disease. The mycobacteria isolated were M. intracellulare (n = 9, 15.5%), M. fortuitum (n = 8, 13.8%), M. gordonae (n = 7, 12.1%), M. kansasii (n = 6, 10.3%), M. chelonae (n = 5, 8.6%), M. avium (n = 4, 6.9%), M. abscessus (n = 3, 5.2%), M. peregrinum (n = 3, 5.2%), M. triplex (n = 3, 5.2%), M. spp. (n = 3, 5.2%), M. szulgai (n = 3, 5.2%), M. mucogenicum (n = 2, 3.4%), M. lentiflavum (n = 1, 1.7%) and M. simiae (n = 1, 1.7%). The present high percentage of NTM strains among mycobacterial isolates (approximately 11%), suggests the emergence of NTM infections as a public health problem. Larger, multicentre and multiregional studies or mandatory reporting will be required to better understand the changing epidemiology of NTM in patients with lung disease.