Gregory G Anderson, Katharine Segars, Anastacia M Sanchez, Jon W Weeks, Shanil P Haugen, Ruchi Pandey
Introduction. Simethicone is an over-the-counter product that is frequently used by clinicians during endoscopic procedures to reduce foaming and improve visualization. Published studies have found simethicone residue on endoscopes after cleaning and disinfecting the devices as per the manufacturer's instructions. Some literature suggests that simethicone residue may reduce disinfection efficacy and increase the risk of patient infections.Gap Statement. However, there appears to be a lack of direct evidence in the literature to either disprove this or correlate simethicone presence with an increased microbial risk.Aim: Research was conducted to evaluate the in vitro impact of simethicone on disinfection efficacy.Methodology. Bacteria were grown in a microtitre plate assay in the presence of a range of simethicone concentrations and then treated with a disinfectant. Bacterial growth was assessed by spotting each microtitre well onto an agar plate.Results. The results demonstrated that, under the conditions tested, simethicone did not reduce the efficacy of Cidex ortho-phthalaldehyde disinfectant, which demonstrated at least a 6-log unit reduction in bacterial viability. Additional experiments showed that direct exposure to 66 mg ml-1 of simethicone reduced bacterial viability.Conclusion. These results indicate that simethicone may not reduce the bactericidal efficacy of disinfectant during reprocessing, under certain conditions.
{"title":"Effect of simethicone on the bactericidal efficacy of a high-level disinfectant.","authors":"Gregory G Anderson, Katharine Segars, Anastacia M Sanchez, Jon W Weeks, Shanil P Haugen, Ruchi Pandey","doi":"10.1099/jmm.0.001902","DOIUrl":"https://doi.org/10.1099/jmm.0.001902","url":null,"abstract":"<p><p><b>Introduction.</b> Simethicone is an over-the-counter product that is frequently used by clinicians during endoscopic procedures to reduce foaming and improve visualization. Published studies have found simethicone residue on endoscopes after cleaning and disinfecting the devices as per the manufacturer's instructions. Some literature suggests that simethicone residue may reduce disinfection efficacy and increase the risk of patient infections.<b>Gap Statement.</b> However, there appears to be a lack of direct evidence in the literature to either disprove this or correlate simethicone presence with an increased microbial risk.<b>Aim</b>: Research was conducted to evaluate the <i>in vitro</i> impact of simethicone on disinfection efficacy.<b>Methodology.</b> Bacteria were grown in a microtitre plate assay in the presence of a range of simethicone concentrations and then treated with a disinfectant. Bacterial growth was assessed by spotting each microtitre well onto an agar plate.<b>Results.</b> The results demonstrated that, under the conditions tested, simethicone did not reduce the efficacy of Cidex ortho-phthalaldehyde disinfectant, which demonstrated at least a 6-log unit reduction in bacterial viability. Additional experiments showed that direct exposure to 66 mg ml<sup>-1</sup> of simethicone reduced bacterial viability.<b>Conclusion.</b> These results indicate that simethicone may not reduce the bactericidal efficacy of disinfectant during reprocessing, under certain conditions.</p>","PeriodicalId":94093,"journal":{"name":"Journal of medical microbiology","volume":"73 10","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142373942","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Huixia Chen, Liqin Zhao, Yan Liang, Hui Xie, Siyu Chen, Lin Wang, Xiao Han
Introduction. Negative changes in the microbial composition have been extensively studied in individuals with periodontal disease.Gap Statement. The changes in the oral microbiota after treating this disease are still unknown.Aim. We sought to elucidate the distinctive traits of salivary microbiota in individuals displaying healthy gums and those with severe periodontitis (SP) and examine the influence of periodontal therapy.Methodology. Periodontal pocket depths were examined to determine disease severity. The presence and quantity of oral Helicobacter pylori (associated with periodontal disease) were determined. Sequencing of 16S ribosomal DNA and bioinformatic analyses were performed to assess oral bacterial compositions in patients.Results. Sequencing analysis of 16S ribosomal DNA revealed a significant reduction in the abundance of operational taxonomic units (OTUs) and Chao1 and Abundance coverage-based estimator(ACE) indices in the oral cavities of individuals with SP compared to those of the healthy controls. However, these parameters showed significant recovery after appropriate treatment severe periodontitis after treatment (TSP). Additionally, the levels of harmful Bacillales and Spirochetes significantly increased, whereas the presence of beneficial Euryarchaeota significantly decreased in the SP group. The TSP group exhibited considerably augmented abundances of Burkholderiaceae and Veillonella, while noteworthy reductions in the pathogenic microbiota (Clostridia, Fusobacteria and Spirochaetes) were noticed compared to those of the SP group. Functionally, these modified OTUs were extensively implicated in 41 metabolic pathways.Conclusion. Our study demonstrates that nonsurgical periodontal therapy can effectively reduce the diversity of the oral microbiota, thereby potentially enhancing the treatment efficacy in patients with periodontal disease.
简介。人们对牙周病患者体内微生物组成的负面变化进行了广泛研究。治疗牙周病后口腔微生物群的变化仍然未知。我们试图阐明健康牙龈和严重牙周炎(SP)患者唾液微生物群的独特特征,并研究牙周治疗的影响。检查牙周袋深度以确定疾病严重程度。测定口腔幽门螺旋杆菌(与牙周病有关)的存在和数量。进行 16S 核糖体 DNA 测序和生物信息学分析,以评估患者的口腔细菌组成。16S 核糖体 DNA 测序分析表明,与健康对照组相比,SP 患者口腔中操作分类单元(OTU)的丰度以及 Chao1 和基于丰度覆盖率的估算器(ACE)指数显著降低。然而,经过适当治疗后,这些参数在严重牙周炎治疗后(TSP)有明显恢复。此外,在 SP 组中,有害的芽孢杆菌和螺旋体的含量明显增加,而有益的真菌则明显减少。与 SP 组相比,TSP 组中 Burkholderiaceae 和 Veillonella 的数量明显增加,而致病微生物群(梭菌、镰刀菌和螺旋体)则明显减少。从功能上看,这些改变的OTU广泛参与了41种代谢途径。我们的研究表明,非手术牙周治疗可有效减少口腔微生物群的多样性,从而提高牙周病患者的治疗效果。
{"title":"Impact of periodontal therapy on oral bacterial composition in individuals diagnosed with advanced periodontal disease.","authors":"Huixia Chen, Liqin Zhao, Yan Liang, Hui Xie, Siyu Chen, Lin Wang, Xiao Han","doi":"10.1099/jmm.0.001913","DOIUrl":"https://doi.org/10.1099/jmm.0.001913","url":null,"abstract":"<p><p><b>Introduction.</b> Negative changes in the microbial composition have been extensively studied in individuals with periodontal disease.<b>Gap Statement.</b> The changes in the oral microbiota after treating this disease are still unknown.<b>Aim.</b> We sought to elucidate the distinctive traits of salivary microbiota in individuals displaying healthy gums and those with severe periodontitis (SP) and examine the influence of periodontal therapy.<b>Methodology.</b> Periodontal pocket depths were examined to determine disease severity. The presence and quantity of oral <i>Helicobacter pylori</i> (associated with periodontal disease) were determined. Sequencing of 16S ribosomal DNA and bioinformatic analyses were performed to assess oral bacterial compositions in patients.<b>Results.</b> Sequencing analysis of 16S ribosomal DNA revealed a significant reduction in the abundance of operational taxonomic units (OTUs) and Chao1 and Abundance coverage-based estimator(ACE) indices in the oral cavities of individuals with SP compared to those of the healthy controls. However, these parameters showed significant recovery after appropriate treatment severe periodontitis after treatment (TSP). Additionally, the levels of harmful Bacillales and Spirochetes significantly increased, whereas the presence of beneficial <i>Euryarchaeota</i> significantly decreased in the SP group. The TSP group exhibited considerably augmented abundances of Burkholderiaceae and Veillonella, while noteworthy reductions in the pathogenic microbiota (Clostridia, Fusobacteria and Spirochaetes) were noticed compared to those of the SP group. Functionally, these modified OTUs were extensively implicated in 41 metabolic pathways.<b>Conclusion.</b> Our study demonstrates that nonsurgical periodontal therapy can effectively reduce the diversity of the oral microbiota, thereby potentially enhancing the treatment efficacy in patients with periodontal disease.</p>","PeriodicalId":94093,"journal":{"name":"Journal of medical microbiology","volume":"73 10","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142484391","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Carmen A Contreras, Tracy H Hazen, Carmen Guadarrama, Ramón Cervantes-Rivera, Theresa J Ochoa, Pablo Vinuesa, David A Rasko, Jose L Puente
Introduction. Enteropathogenic Escherichia coli (EPEC) strains pose a significant threat as a leading cause of severe childhood diarrhoea in developing nations. EPEC pathogenicity relies on the type III secretion system (T3SS) encoded by the locus of enterocyte effacement (LEE), facilitating the secretion and translocation of bacterial effector proteins.Gap Statement. While the regulatory roles of PerC (plasmid-encoded regulator) and GrlA (global regulator of LEE-activator) in ler expression and LEE gene activation are well-documented in the EPEC prototype strain E2348/69, understanding the variability in LEE gene expression control mechanisms among clinical EPEC isolates remains an area requiring further investigation.Aim. This study aims to explore the diversity in LEE gene expression control mechanisms among clinical EPEC isolates through a comparative analysis of secretion profiles under defined growth conditions favouring either PerC- or GrlA-mediated activation of LEE expression.Methodology. We compared T3SS-dependent secretion patterns and promoter expression in both typical EPEC (tEPEC) and atypical EPEC (aEPEC) clinical isolates under growth conditions favouring either PerC- or GrlA-mediated activation of LEE expression. Additionally, we conducted promoter reporter activity assays, quantitative real-time PCR and Western blot experiments to assess gene expression activity.Results. Significant differences in T3SS-dependent secretion were observed among tEPEC and aEPEC strains, independent of LEE sequence variations or T3SS gene functionality. Notably, a clinical tEPEC isolate exhibited increased secretion levels under repressive growth conditions and in the absence of both PerC and GrlA, implicating an alternative mechanism in the activation of Ler (LEE-encoded regulator) expression.Conclusion. Our findings indicate that uncharacterized LEE regulatory mechanisms contribute to phenotypic diversity among clinical EPEC isolates, though their impact on clinical outcomes remains unknown. This challenges the conventional understanding based on reference strains and highlights the need to investigate beyond established models to comprehensively elucidate EPEC pathogenesis.
导言。肠致病性大肠杆菌(EPEC)菌株是发展中国家儿童严重腹泻的主要病因,对儿童构成了严重威胁。EPEC 的致病性依赖于由肠道细胞损伤基因座(LEE)编码的 III 型分泌系统(T3SS),该系统可促进细菌效应蛋白的分泌和转运。虽然在 EPEC 原型菌株 E2348/69 中,PerC(质粒编码的调节因子)和 GrlA(LEE 激活因子的全局调节因子)在 ler 表达和 LEE 基因激活中的调控作用已得到充分证实,但了解临床 EPEC 分离株之间 LEE 基因表达调控机制的变异性仍是一个需要进一步研究的领域。本研究旨在通过比较分析在有利于PerC或GrlA介导的LEE表达激活的特定生长条件下的分泌谱,探索临床EPEC分离株之间LEE基因表达控制机制的多样性。我们比较了典型 EPEC(tEPEC)和非典型 EPEC(aEPEC)临床分离株在有利于 PerC 或 GrlA 介导的激活 LEE 表达的生长条件下依赖 T3SS 的分泌模式和启动子表达。此外,我们还进行了启动子报告活性测定、定量实时 PCR 和 Western 印迹实验,以评估基因表达活性。在 tEPEC 和 aEPEC 菌株之间观察到了 T3SS 依赖性分泌的显著差异,这与 LEE 序列变异或 T3SS 基因的功能无关。值得注意的是,一种临床 tEPEC 分离物在抑制性生长条件下以及在 PerC 和 GrlA 均缺失的情况下表现出分泌水平升高,这表明 Ler(LEE 编码的调节因子)表达的激活存在另一种机制。我们的研究结果表明,未表征的 LEE 调控机制导致了临床 EPEC 分离物的表型多样性,但它们对临床结果的影响仍然未知。这对基于参考菌株的传统认识提出了挑战,并凸显了超越既有模型进行研究以全面阐明 EPEC 发病机制的必要性。
{"title":"Phenotypic diversity of type III secretion system activity in enteropathogenic <i>Escherichia coli</i> clinical isolates.","authors":"Carmen A Contreras, Tracy H Hazen, Carmen Guadarrama, Ramón Cervantes-Rivera, Theresa J Ochoa, Pablo Vinuesa, David A Rasko, Jose L Puente","doi":"10.1099/jmm.0.001907","DOIUrl":"10.1099/jmm.0.001907","url":null,"abstract":"<p><p><b>Introduction.</b> Enteropathogenic <i>Escherichia coli</i> (EPEC) strains pose a significant threat as a leading cause of severe childhood diarrhoea in developing nations. EPEC pathogenicity relies on the type III secretion system (T3SS) encoded by the locus of enterocyte effacement (LEE), facilitating the secretion and translocation of bacterial effector proteins.<b>Gap Statement.</b> While the regulatory roles of PerC (plasmid-encoded regulator) and GrlA (global regulator of LEE-activator) in <i>ler</i> expression and LEE gene activation are well-documented in the EPEC prototype strain E2348/69, understanding the variability in LEE gene expression control mechanisms among clinical EPEC isolates remains an area requiring further investigation.<b>Aim.</b> This study aims to explore the diversity in LEE gene expression control mechanisms among clinical EPEC isolates through a comparative analysis of secretion profiles under defined growth conditions favouring either PerC- or GrlA-mediated activation of LEE expression.<b>Methodology.</b> We compared T3SS-dependent secretion patterns and promoter expression in both typical EPEC (tEPEC) and atypical EPEC (aEPEC) clinical isolates under growth conditions favouring either PerC- or GrlA-mediated activation of LEE expression. Additionally, we conducted promoter reporter activity assays, quantitative real-time PCR and Western blot experiments to assess gene expression activity.<b>Results.</b> Significant differences in T3SS-dependent secretion were observed among tEPEC and aEPEC strains, independent of LEE sequence variations or T3SS gene functionality. Notably, a clinical tEPEC isolate exhibited increased secretion levels under repressive growth conditions and in the absence of both PerC and GrlA, implicating an alternative mechanism in the activation of Ler (LEE-encoded regulator) expression.<b>Conclusion.</b> Our findings indicate that uncharacterized LEE regulatory mechanisms contribute to phenotypic diversity among clinical EPEC isolates, though their impact on clinical outcomes remains unknown. This challenges the conventional understanding based on reference strains and highlights the need to investigate beyond established models to comprehensively elucidate EPEC pathogenesis.</p>","PeriodicalId":94093,"journal":{"name":"Journal of medical microbiology","volume":"73 10","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11493143/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142484392","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Klaudia Bartoszewicz, Mateusz Bartoszewicz, Samuel Stróż, Anna Stasiak-Barmuta, Piotr Kosiorek
Introduction. The Coronavirus Disease 2019 (COVID-19) pandemic has had a significant impact on global healthcare, with high mortality and severe complications remaining a major concern. Understanding the predictors of COVID-19 severity may improve patient management and outcomes. While considerable research has focused on the pathogenesis of the virus and vaccine development, the identification of reliable demographic, clinical and laboratory predictors of severe disease remains critical.Hypothesis. Specific demographic factors, clinical signs and laboratory markers can reliably predict the severity of COVID-19. A comprehensive analysis integrating these predictors could provide a more accurate prognosis and guide timely interventions.Aim. The aim of this study is to identify and evaluate the demographic, clinical and laboratory factors that can serve as reliable predictors of severe COVID-19, thereby aiding in the prediction and prevention of adverse outcomes.Methodology. The methods of analysis, synthesis, generalization and descriptive statistics were used to achieve this objective.Results. The analysis showed that demographic factors such as age over 60 and male sex are significant predictors of severe COVID-19. Clinical predictors include respiratory symptoms, especially dyspnoea, and comorbidities such as hypertension, coronary artery disease, chronic obstructive pulmonary disease, respiratory failure, asthma, diabetes mellitus and obesity. Laboratory markers with high prognostic value include elevated levels of C-reactive protein, interleukin-6, ferritin, neutrophil/lymphocyte ratio, d-dimer, aspartate aminotransferase enzyme and decreased lymphocyte count.Conclusion. The study concludes that a holistic approach incorporating demographic, clinical and laboratory data is essential to accurately predict the severity of COVID-19. This integrated model may significantly improve patient prognosis by facilitating early identification of high-risk individuals and allowing timely, targeted interventions. The results highlight the importance of comprehensive patient assessment in managing and mitigating the impact of COVID-19.
{"title":"Predictors of the severity of the course of COVID-19: demographic factors, clinical signs and laboratory markers.","authors":"Klaudia Bartoszewicz, Mateusz Bartoszewicz, Samuel Stróż, Anna Stasiak-Barmuta, Piotr Kosiorek","doi":"10.1099/jmm.0.001911","DOIUrl":"10.1099/jmm.0.001911","url":null,"abstract":"<p><p><b>Introduction.</b> The Coronavirus Disease 2019 (COVID-19) pandemic has had a significant impact on global healthcare, with high mortality and severe complications remaining a major concern. Understanding the predictors of COVID-19 severity may improve patient management and outcomes. While considerable research has focused on the pathogenesis of the virus and vaccine development, the identification of reliable demographic, clinical and laboratory predictors of severe disease remains critical.<b>Hypothesis.</b> Specific demographic factors, clinical signs and laboratory markers can reliably predict the severity of COVID-19. A comprehensive analysis integrating these predictors could provide a more accurate prognosis and guide timely interventions.<b>Aim.</b> The aim of this study is to identify and evaluate the demographic, clinical and laboratory factors that can serve as reliable predictors of severe COVID-19, thereby aiding in the prediction and prevention of adverse outcomes.<b>Methodology.</b> The methods of analysis, synthesis, generalization and descriptive statistics were used to achieve this objective.<b>Results.</b> The analysis showed that demographic factors such as age over 60 and male sex are significant predictors of severe COVID-19. Clinical predictors include respiratory symptoms, especially dyspnoea, and comorbidities such as hypertension, coronary artery disease, chronic obstructive pulmonary disease, respiratory failure, asthma, diabetes mellitus and obesity. Laboratory markers with high prognostic value include elevated levels of C-reactive protein, interleukin-6, ferritin, neutrophil/lymphocyte ratio, d-dimer, aspartate aminotransferase enzyme and decreased lymphocyte count.<b>Conclusion.</b> The study concludes that a holistic approach incorporating demographic, clinical and laboratory data is essential to accurately predict the severity of COVID-19. This integrated model may significantly improve patient prognosis by facilitating early identification of high-risk individuals and allowing timely, targeted interventions. The results highlight the importance of comprehensive patient assessment in managing and mitigating the impact of COVID-19.</p>","PeriodicalId":94093,"journal":{"name":"Journal of medical microbiology","volume":"73 10","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142396388","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Annamaria Porreca, Eliana Ibrahimi, Fabrizio Maturo, Laura Judith Marcos Zambrano, Melisa Meto, Marta B Lopes
Introduction. The study addresses the challenge of utilizing human gut microbiome data for the early detection of colorectal cancer (CRC). The research emphasizes the potential of using machine learning techniques to analyze complex microbiome datasets, providing a non-invasive approach to identifying CRC-related microbial markers.Hypothesis/Gap Statement. The primary hypothesis is that a robust machine learning-based analysis of 16S rRNA microbiome data can identify specific microbial features that serve as effective biomarkers for CRC detection, overcoming the limitations of classical statistical models in high-dimensional settings.Aim. The primary objective of this study is to explore and validate the potential of the human microbiome, specifically in the colon, as a valuable source of biomarkers for colorectal cancer (CRC) detection and progression. The focus is on developing a classifier that effectively predicts the presence of CRC and normal samples based on the analysis of three previously published faecal 16S rRNA sequencing datasets.Methodology. To achieve the aim, various machine learning techniques are employed, including random forest (RF), recursive feature elimination (RFE) and a robust correlation-based technique known as the fuzzy forest (FF). The study utilizes these methods to analyse the three datasets, comparing their performance in predicting CRC and normal samples. The emphasis is on identifying the most relevant microbial features (taxa) associated with CRC development via partial dependence plots, i.e. a machine learning tool focused on explainability, visualizing how a feature influences the predicted outcome.Results. The analysis of the three faecal 16S rRNA sequencing datasets reveals the consistent and superior predictive performance of the FF compared to the RF and RFE. Notably, FF proves effective in addressing the correlation problem when assessing the importance of microbial taxa in explaining the development of CRC. The results highlight the potential of the human microbiome as a non-invasive means to detect CRC and underscore the significance of employing FF for improved predictive accuracy.Conclusion. In conclusion, this study underscores the limitations of classical statistical techniques in handling high-dimensional information such as human microbiome data. The research demonstrates the potential of the human microbiome, specifically in the colon, as a valuable source of biomarkers for CRC detection. Applying machine learning techniques, particularly the FF, is a promising approach for building a classifier to predict CRC and normal samples. The findings advocate for integrating FF to overcome the challenges associated with correlation when identifying crucial microbial features linked to CRC development.
{"title":"Robust prediction of colorectal cancer via gut microbiome 16S rRNA sequencing data.","authors":"Annamaria Porreca, Eliana Ibrahimi, Fabrizio Maturo, Laura Judith Marcos Zambrano, Melisa Meto, Marta B Lopes","doi":"10.1099/jmm.0.001903","DOIUrl":"https://doi.org/10.1099/jmm.0.001903","url":null,"abstract":"<p><p><b>Introduction.</b> The study addresses the challenge of utilizing human gut microbiome data for the early detection of colorectal cancer (CRC). The research emphasizes the potential of using machine learning techniques to analyze complex microbiome datasets, providing a non-invasive approach to identifying CRC-related microbial markers.<b>Hypothesis/Gap Statement.</b> The primary hypothesis is that a robust machine learning-based analysis of 16S rRNA microbiome data can identify specific microbial features that serve as effective biomarkers for CRC detection, overcoming the limitations of classical statistical models in high-dimensional settings.<b>Aim.</b> The primary objective of this study is to explore and validate the potential of the human microbiome, specifically in the colon, as a valuable source of biomarkers for colorectal cancer (CRC) detection and progression. The focus is on developing a classifier that effectively predicts the presence of CRC and normal samples based on the analysis of three previously published faecal 16S rRNA sequencing datasets.<b>Methodology.</b> To achieve the aim, various machine learning techniques are employed, including random forest (RF), recursive feature elimination (RFE) and a robust correlation-based technique known as the fuzzy forest (FF). The study utilizes these methods to analyse the three datasets, comparing their performance in predicting CRC and normal samples. The emphasis is on identifying the most relevant microbial features (taxa) associated with CRC development via partial dependence plots, i.e. a machine learning tool focused on explainability, visualizing how a feature influences the predicted outcome.<b>Results.</b> The analysis of the three faecal 16S rRNA sequencing datasets reveals the consistent and superior predictive performance of the FF compared to the RF and RFE. Notably, FF proves effective in addressing the correlation problem when assessing the importance of microbial taxa in explaining the development of CRC. The results highlight the potential of the human microbiome as a non-invasive means to detect CRC and underscore the significance of employing FF for improved predictive accuracy.<b>Conclusion.</b> In conclusion, this study underscores the limitations of classical statistical techniques in handling high-dimensional information such as human microbiome data. The research demonstrates the potential of the human microbiome, specifically in the colon, as a valuable source of biomarkers for CRC detection. Applying machine learning techniques, particularly the FF, is a promising approach for building a classifier to predict CRC and normal samples. The findings advocate for integrating FF to overcome the challenges associated with correlation when identifying crucial microbial features linked to CRC development.</p>","PeriodicalId":94093,"journal":{"name":"Journal of medical microbiology","volume":"73 10","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142396389","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mehmood Qadir, Muhammad Tahir Khan, Sajjad Ahmed Khan, Muhammad Akram, Julio Ortiz Canseco, Rani Faryal, Dong Qing Wei, Sabira Tahseen
Introduction. The discordance between phenotypic and molecular methods of rifampicin (RIF) drug susceptibility testing (DST) in Mycobacterium tuberculosis poses a significant challenge, potentially resulting in misdiagnosis and inappropriate treatment.Hypothesis/gap statement. A comparison of RIF phenotypic and molecular methods for DST, including whole genome sequencing (WGS), may provide a better understanding of resistance mechanisms.Aim. This study aims to compare RIF DST in M. tuberculosis using two phenotypic and molecular methods including the GeneXpert RIF Assay (GX) and WGS for better understanding.Methodology. The study evaluated two phenotypic liquid medium methods [Lowenstein-Jensen (LJ) and Mycobacterium Growth Indicator Tube (MGIT)], one targeted molecular method (GX), and one WGS method. Moreover, mutational frequency in ponA1 and ponA2 was also screened in the current and previous RIF resistance M. tuberculosis genomic isolates to find their compensatory role.Results. A total of 25 RIF-resistant isolates, including nine from treatment failures and relapse cases with both discordant and concordant DST results on LJ, MGIT and GX, were subjected to WGS. The phenotypic DST results indicated that 11 isolates (44%) were susceptible on LJ and MGIT but resistant on GX. These isolates exhibited multiple mutations in rpoB, including Thr444>Ala, Leu430>Pro, Leu430>Arg, Asp435>Gly, His445>Asn and Asn438>Lys. Conversely, four isolates that were susceptible on GX and MGIT but resistant on LJ were wild type for rpoB in WGS. However, these isolates possessed several novel mutations in the PonA1 gene, including a 10 nt insertion and two nonsynonymous mutations (Ala394>Ser, Pro631>Ser), as well as one nonsynonymous mutation (Pro780>Arg) in PonA2. The discordance rate of RIF DST is higher on MGIT than on LJ and GX when compared to WGS. These discordances in the Delhi/CAS lineages were primarily associated with failure and relapse cases.Conclusion. The WGS of RIF resistance is relatively expensive, but it may be considered for isolates with discordant DST results on MGIT, LJ and GX to ensure accurate diagnosis and appropriate treatment options.
{"title":"Unveiling the complexity of rifampicin drug susceptibility testing in <i>Mycobacterium tuberculosis</i>: comparative analysis with next-generation sequencing.","authors":"Mehmood Qadir, Muhammad Tahir Khan, Sajjad Ahmed Khan, Muhammad Akram, Julio Ortiz Canseco, Rani Faryal, Dong Qing Wei, Sabira Tahseen","doi":"10.1099/jmm.0.001884","DOIUrl":"https://doi.org/10.1099/jmm.0.001884","url":null,"abstract":"<p><p><b>Introduction.</b> The discordance between phenotypic and molecular methods of rifampicin (RIF) drug susceptibility testing (DST) in <i>Mycobacterium tuberculosis</i> poses a significant challenge, potentially resulting in misdiagnosis and inappropriate treatment.<b>Hypothesis/gap statement.</b> A comparison of RIF phenotypic and molecular methods for DST, including whole genome sequencing (WGS), may provide a better understanding of resistance mechanisms.<b>Aim.</b> This study aims to compare RIF DST in <i>M. tuberculosis</i> using two phenotypic and molecular methods including the GeneXpert RIF Assay (GX) and WGS for better understanding.<b>Methodology.</b> The study evaluated two phenotypic liquid medium methods [Lowenstein-Jensen (LJ) and Mycobacterium Growth Indicator Tube (MGIT)], one targeted molecular method (GX), and one WGS method. Moreover, mutational frequency in <i>ponA1</i> and <i>ponA2</i> was also screened in the current and previous RIF resistance <i>M. tuberculosis</i> genomic isolates to find their compensatory role.<b>Results.</b> A total of 25 RIF-resistant isolates, including nine from treatment failures and relapse cases with both discordant and concordant DST results on LJ, MGIT and GX, were subjected to WGS. The phenotypic DST results indicated that 11 isolates (44%) were susceptible on LJ and MGIT but resistant on GX. These isolates exhibited multiple mutations in <i>rpoB</i>, including Thr444>Ala, Leu430>Pro, Leu430>Arg, Asp435>Gly, His445>Asn and Asn438>Lys. Conversely, four isolates that were susceptible on GX and MGIT but resistant on LJ were wild type for <i>rpoB</i> in WGS. However, these isolates possessed several novel mutations in the PonA1 gene, including a 10 nt insertion and two nonsynonymous mutations (Ala394>Ser, Pro631>Ser), as well as one nonsynonymous mutation (Pro780>Arg) in PonA2. The discordance rate of RIF DST is higher on MGIT than on LJ and GX when compared to WGS. These discordances in the Delhi/CAS lineages were primarily associated with failure and relapse cases.<b>Conclusion.</b> The WGS of RIF resistance is relatively expensive, but it may be considered for isolates with discordant DST results on MGIT, LJ and GX to ensure accurate diagnosis and appropriate treatment options.</p>","PeriodicalId":94093,"journal":{"name":"Journal of medical microbiology","volume":"73 9","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142127766","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hafize Dilşad Yanık, Nefise Akçelik, Elif Gamze Has, Mustafa Akçelik
Introduction. Disruptions in gut microbiota, known as dysbiosis, have been increasingly linked to pathogenic infections, with Salmonella Typhimurium being a notable contributor to these disturbances.Hypothesis. We hypothesize that the S. Typhimurium 14028 WT strain induces significant dysbiosis in the rat gut microbiota and that the dam and seqA genes play crucial roles in this process.Aim. In this study, it was aimed at investigating the dysbiotic activity of the S. Typhimurium 14028 WT strain on the rat gut microbiota and the roles of dam and seqA genes on this activity.Method. Changes in the rat gut microbiota were determined by examining the anal swap samples taken from the experimental groups of these animals using 16S rRNA high-throughput sequencing technology.Results. In the experimental groups, the dominant phyla were determined to be Firmicutes and Bacteroidetes (P<0.05). However, while the rate of Bacteroidetes was significantly reduced in those treated with the WT and seqA mutants, no significant difference was observed in the dam mutant compared to the control group (P<0.05). In all experimental animals, the dominant species was determined to be Prevotella copri, regardless of the experiment time and application. The analysis results of the samples taken on the third day from the rat groups infected with the S. Typhimurium 14028 WT strain (W2) presented the most striking data of this study.Conclusion. Through distance analysis, we demonstrated that a successful Salmonella infection completely changes the composition of the microbiota, dramatically reduces species diversity and richness in the microbiota and encourages the growth of opportunistic pathogens.
导言。肠道微生物群紊乱(称为菌群失调)与致病性感染的关系日益密切,而伤寒沙门氏菌是造成这些紊乱的一个显著因素。我们假设伤寒沙门氏菌 14028 WT 菌株会诱导大鼠肠道微生物群发生严重的菌群失调,而 Dam 和 seqA 基因在这一过程中起着至关重要的作用。本研究旨在探讨鼠伤寒杆菌 14028 WT 菌株对大鼠肠道微生物群的菌群失调活性以及 dam 和 seqA 基因在这一活性中的作用。采用 16S rRNA 高通量测序技术检测实验组大鼠的肛门交换样本,以确定大鼠肠道微生物群的变化。在实验组中,主要的菌门是固着菌门和类杆菌门(PBacteroidetes),在使用 WT 和 seqA 突变体的实验组中,类杆菌门显著减少,而 Dam 突变体与对照组(PPrevotella copri)相比,无论实验时间和应用情况如何,都没有观察到显著差异。本研究中最引人注目的数据是对感染鼠伤寒杆菌 14028 WT 株(W2)的大鼠组在第三天采集的样本进行的分析结果。通过距离分析,我们证明成功感染沙门氏菌会完全改变微生物群的组成,显著降低微生物群的物种多样性和丰富度,并促进机会性病原体的生长。
{"title":"Relationship of <i>Salmonella</i> Typhimurium 14028 strain and its <i>dam</i> and <i>seqA</i> mutants with gut microbiota dysbiosis in rats.","authors":"Hafize Dilşad Yanık, Nefise Akçelik, Elif Gamze Has, Mustafa Akçelik","doi":"10.1099/jmm.0.001893","DOIUrl":"https://doi.org/10.1099/jmm.0.001893","url":null,"abstract":"<p><p><b>Introduction.</b> Disruptions in gut microbiota, known as dysbiosis, have been increasingly linked to pathogenic infections, with <i>Salmonella</i> Typhimurium being a notable contributor to these disturbances.<b>Hypothesis.</b> We hypothesize that the <i>S</i>. Typhimurium 14028 WT strain induces significant dysbiosis in the rat gut microbiota and that the <i>dam</i> and <i>seqA</i> genes play crucial roles in this process.<b>Aim</b>. In this study, it was aimed at investigating the dysbiotic activity of the <i>S</i>. Typhimurium 14028 WT strain on the rat gut microbiota and the roles of <i>dam</i> and <i>seqA</i> genes on this activity.<b>Method.</b> Changes in the rat gut microbiota were determined by examining the anal swap samples taken from the experimental groups of these animals using 16S rRNA high-throughput sequencing technology.<b>Results.</b> In the experimental groups, the dominant phyla were determined to be <i>Firmicutes</i> and <i>Bacteroidetes</i> (<i>P</i><0.05). However, while the rate of <i>Bacteroidetes</i> was significantly reduced in those treated with the WT and <i>seqA</i> mutants, no significant difference was observed in the <i>dam</i> mutant compared to the control group (<i>P</i><0.05). In all experimental animals, the dominant species was determined to be <i>Prevotella copri</i>, regardless of the experiment time and application. The analysis results of the samples taken on the third day from the rat groups infected with the <i>S.</i> Typhimurium 14028 WT strain (W2) presented the most striking data of this study.<b>Conclusion.</b> Through distance analysis, we demonstrated that a successful <i>Salmonella</i> infection completely changes the composition of the microbiota, dramatically reduces species diversity and richness in the microbiota and encourages the growth of opportunistic pathogens.</p>","PeriodicalId":94093,"journal":{"name":"Journal of medical microbiology","volume":"73 9","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142335312","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Leah Rankine-Wilson, Teresa Oncken, Irshan Basrewan, Courtney Jeffery, Todd M Pryce, Rebecca Wake, Aus A L Molan, T F Paton, Tim J J Inglis
Background. The COVID-19 pandemic demonstrated a need for robust SARS-CoV-2 test evaluation infrastructure to underpin biosecurity and protect the population during a pandemic health emergency.Gap statement. The first generation of rapid antigen tests was less accurate than molecular methods due to their inherent sensitivity and specificity shortfalls, compounded by the consequences of self-testing. This created a need for more accurate point-of-care SARS-CoV-2 detection methods.Aim. Here we present the lessons-learned during the COVID-19 emergency response in Western Australia including the detailed set-up, evaluation and operation of rapid antigen test in a state-run drive-through sample collection service during the COVID-19 pandemic after the strict border shutdown ended.Methods. We report a conformity assessment of a novel, second-generation rapid antigen test (Virulizer) comprising a technician-operated rapid lateral flow immunoassay with fluorescence-based detection.Results. The Virulizer rapid antigen test demonstrated up to 100% sensitivity (95% CI: 61.0-100%), 91.94% specificity (95% CI: 82.5-96.5%) and 92.65% accuracy when compared to a commercial PCR assay method. Wide confidence intervals in our series reflect the limits of small sample size. Nevertheless, the Virulizer assay performance was well-suited to point-of-care screening for SARS-CoV-2 in a drive-through clinic setting.Conclusion. The adaptive evaluation process necessary under changing pandemic conditions enabled assessment of a simple sample collection and point-of-care testing process, and showed how this system could be rapidly deployed for SARS-CoV-2 testing, including to regional and remote settings.
{"title":"Lessons learned: drive-through COVID-19 clinic testing during an adaptive epidemic response and a point-of-care test assessment of a computer-read rapid lateral flow immunoassay with fluorescence-based detection.","authors":"Leah Rankine-Wilson, Teresa Oncken, Irshan Basrewan, Courtney Jeffery, Todd M Pryce, Rebecca Wake, Aus A L Molan, T F Paton, Tim J J Inglis","doi":"10.1099/jmm.0.001875","DOIUrl":"10.1099/jmm.0.001875","url":null,"abstract":"<p><p><b>Background.</b> The COVID-19 pandemic demonstrated a need for robust SARS-CoV-2 test evaluation infrastructure to underpin biosecurity and protect the population during a pandemic health emergency.<b>Gap statement.</b> The first generation of rapid antigen tests was less accurate than molecular methods due to their inherent sensitivity and specificity shortfalls, compounded by the consequences of self-testing. This created a need for more accurate point-of-care SARS-CoV-2 detection methods.<b>Aim.</b> Here we present the lessons-learned during the COVID-19 emergency response in Western Australia including the detailed set-up, evaluation and operation of rapid antigen test in a state-run drive-through sample collection service during the COVID-19 pandemic after the strict border shutdown ended.<b>Methods.</b> We report a conformity assessment of a novel, second-generation rapid antigen test (Virulizer) comprising a technician-operated rapid lateral flow immunoassay with fluorescence-based detection.<b>Results.</b> The Virulizer rapid antigen test demonstrated up to 100% sensitivity (95% CI: 61.0-100%), 91.94% specificity (95% CI: 82.5-96.5%) and 92.65% accuracy when compared to a commercial PCR assay method. Wide confidence intervals in our series reflect the limits of small sample size. Nevertheless, the Virulizer assay performance was well-suited to point-of-care screening for SARS-CoV-2 in a drive-through clinic setting.<b>Conclusion.</b> The adaptive evaluation process necessary under changing pandemic conditions enabled assessment of a simple sample collection and point-of-care testing process, and showed how this system could be rapidly deployed for SARS-CoV-2 testing, including to regional and remote settings.</p>","PeriodicalId":94093,"journal":{"name":"Journal of medical microbiology","volume":"73 9","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142116600","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Introduction. Hepatocellular carcinoma (HCC) is one of the deadliest cancers worldwide.Gap statement. Monitoring of HCC and predicting its immunotherapy responses are challenging.Aim. This study explored the potential of the gut microbiome for HCC monitoring and predicting HCC immunotherapy responses.Methods. DNA samples were collected from the faeces of 22 patients with HCC treated with atezolizumab/bevacizumab (Atz/Bev) and 85 healthy controls. The gut microbiome was analysed using 16S rRNA next-generation sequencing and quantitative PCR (qPCR).Results. The microbiomes of patients with HCC demonstrated significant enrichment of Lactobacillus, particularly Lactobacillus fermentum, and Streptococcus, notably Streptococcus anginosus. Comparative analysis between Atz/Bev responders (R) and non-responders (NR) revealed a higher abundance of Bacteroides stercoris in the NR group and Bacteroides coprocola in the R group. Using qPCR analysis, we observed elevated levels of S. anginosus and reduced levels of 5α-reductase genes, essential for the synthesis of isoallolithocholic acid, in HCC patients compared to controls. Additionally, the analysis confirmed a significantly lower abundance of B. stercoris in the Atz/Bev R group relative to the NR group.Conclusions. The gut microbiome analysis and specific gene quantification via qPCR could provide a rapid, less invasive, and cost-effective approach for assessing the increased risk of HCC, monitoring patient status, and predicting immunotherapy responses.
简介:肝细胞癌(HCC)是全球最致命的癌症之一。肝细胞癌(HCC)是全球最致命的癌症之一。监测 HCC 和预测其免疫疗法反应具有挑战性。本研究探讨了肠道微生物组在监测 HCC 和预测 HCC 免疫疗法反应方面的潜力。从22名接受阿特珠单抗/贝伐单抗(Atzolizumab/Bev)治疗的HCC患者和85名健康对照者的粪便中采集DNA样本。使用 16S rRNA 下一代测序和定量 PCR(qPCR)分析肠道微生物组。结果显示,HCC 患者的微生物组中乳酸杆菌(尤其是发酵乳酸杆菌)和链球菌(尤其是副链球菌)明显增多。对 Atz/Bev 反应者(R)和非反应者(NR)的比较分析表明,NR 组中的 Bacteroides stercoris 和 R 组中的 Bacteroides coprocola 含量较高。通过 qPCR 分析,我们观察到与对照组相比,HCC 患者的 S. anginosus 水平升高,而合成异全胆酸所必需的 5α 还原酶基因水平降低。此外,分析还证实,与 NR 组相比,Atz/Bev R 组的 B. stercoris 丰度明显较低。通过 qPCR 进行肠道微生物组分析和特异性基因定量,可为评估 HCC 风险增加、监测患者状态和预测免疫疗法反应提供一种快速、低侵入性和经济有效的方法。
{"title":"Altered intestinal <i>Streptococcus anginosus</i> and 5α-reductase gene levels in patients with hepatocellular carcinoma and elevated <i>Bacteroides stercoris</i> in atezolizumab/bevacizumab non-responders.","authors":"Tadashi Fujii, Teiji Kuzuya, Nobuhiro Kondo, Kohei Funasaka, Eizaburo Ohno, Yoshiki Hirooka, Takumi Tochio","doi":"10.1099/jmm.0.001878","DOIUrl":"10.1099/jmm.0.001878","url":null,"abstract":"<p><p><b>Introduction.</b> Hepatocellular carcinoma (HCC) is one of the deadliest cancers worldwide.<b>Gap statement.</b> Monitoring of HCC and predicting its immunotherapy responses are challenging.<b>Aim.</b> This study explored the potential of the gut microbiome for HCC monitoring and predicting HCC immunotherapy responses.<b>Methods.</b> DNA samples were collected from the faeces of 22 patients with HCC treated with atezolizumab/bevacizumab (Atz/Bev) and 85 healthy controls. The gut microbiome was analysed using 16S rRNA next-generation sequencing and quantitative PCR (qPCR).<b>Results.</b> The microbiomes of patients with HCC demonstrated significant enrichment of <i>Lactobacillus</i>, particularly <i>Lactobacillus fermentum</i>, and <i>Streptococcus</i>, notably <i>Streptococcus anginosus</i>. Comparative analysis between Atz/Bev responders (R) and non-responders (NR) revealed a higher abundance of <i>Bacteroides stercoris</i> in the NR group and <i>Bacteroides coprocola</i> in the R group. Using qPCR analysis, we observed elevated levels of <i>S. anginosus</i> and reduced levels of 5α-reductase genes, essential for the synthesis of isoallolithocholic acid, in HCC patients compared to controls. Additionally, the analysis confirmed a significantly lower abundance of <i>B. stercoris</i> in the Atz/Bev R group relative to the NR group.<b>Conclusions.</b> The gut microbiome analysis and specific gene quantification via qPCR could provide a rapid, less invasive, and cost-effective approach for assessing the increased risk of HCC, monitoring patient status, and predicting immunotherapy responses.</p>","PeriodicalId":94093,"journal":{"name":"Journal of medical microbiology","volume":"73 9","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142142201","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Marginal notes, September 2024: in the antimicrobial resistance hot seat.","authors":"Timothy J J Inglis","doi":"10.1099/jmm.0.001906","DOIUrl":"https://doi.org/10.1099/jmm.0.001906","url":null,"abstract":"","PeriodicalId":94093,"journal":{"name":"Journal of medical microbiology","volume":"73 9","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11423852/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142335311","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}