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Purulent pleurisy caused by Salmonella enterica subspecies arizonae: a case report. 亚利桑那亚种肠沙门氏菌致化脓性胸膜炎1例。
Pub Date : 2025-06-20 eCollection Date: 2025-01-01 DOI: 10.1099/acmi.0.000985.v5
Amine Amri, Youssra Boughalem, Elmostafa Benaissa, Yassine Benlahlou, Mariama Chadli

Background. Salmonellosis most commonly presents clinically as typhoid fever or gastroenteritis. Pleuropulmonary infections due to Salmonella are still rare, even though they have often been described in immunocompromised patients. Case presentation. We report a rare case of purulent pleurisy caused by Salmonella enterica subsp. arizonae, occurring in a 50-year-old female with breast cancer who is currently treated with chemotherapy and radiotherapy along with chronic renal failure requiring haemodialysis, who presented with acute chest pain, dyspnoea and haemodynamic instability. After bacteriological identification of Salmonella enterica subsp. arizonae in pleural fluid, antibiotic susceptibility testing was performed. The patient was then started on a broad-spectrum antibiotic, which successfully improved her condition. Conclusion. Our case highlights the implication of Salmonella enterica subsp. arizonae in purulent pleurisy in an immunocompromised patient. An early diagnosis and a proper antibiotic therapy enabled us to reduce the morbidity and mortality risk in our patient.

背景。沙门氏菌病在临床上最常见的表现是伤寒或肠胃炎。由于沙门氏菌引起的胸膜肺感染仍然很罕见,尽管它们经常被描述为免疫功能低下的患者。例演示。我们报告一个罕见的病例化脓性胸膜炎引起肠炎沙门氏菌亚。亚利桑那州,发生于一名患有乳腺癌的50岁女性,目前正在接受化疗和放疗,并伴有需要血液透析的慢性肾衰竭,表现为急性胸痛、呼吸困难和血流动力学不稳定。经肠道沙门氏菌亚种的细菌学鉴定。在胸膜液中进行亚利桑那球菌药敏试验。然后,病人开始使用广谱抗生素,这成功地改善了她的病情。结论。我们的病例突出了肠道沙门氏菌亚种的含义。免疫功能低下患者脓性胸膜炎中的亚利桑那球菌。早期诊断和适当的抗生素治疗使我们能够降低患者的发病率和死亡率。
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引用次数: 0
Draft genome sequence of Flavobacterium aquidurense strain, isolated from untreated wastewater. 从未经处理的废水中分离的aquidurense黄杆菌菌株的基因组序列草图。
Pub Date : 2025-06-20 eCollection Date: 2025-01-01 DOI: 10.1099/acmi.0.000976.v3
Alexander D H Kingdon, Kara D'Arcy, Anya Breen, Claudia McKeown, Ellie Allman, Priyanka Sharma, Amy McLeman, Adam P Roberts

Here, we report the draft 5.8 Mb genome sequence of a Flavobacterium aquidurense isolate from untreated wastewater in Liverpool, United Kingdom. The reported isolate has the potential to produce both flexirubin and β-carotene pigments, and contains an additional biosynthetic gene cluster for a putative novel β-lactone. The genome also contains a gene for a putative β-lactamase blaJOHN-1 analogue, and there are multiple copies of a putative novel insertion sequence of the IS3 family. This genome adds to a growing resource of Flavobacterium spp. sequencing data which can be utilized to investigate microbial pigment production, antimicrobial resistance genes and mobile genetic elements within this genus.

在这里,我们报道了从英国利物浦未经处理的废水中分离出的aquidurense黄杆菌的5.8 Mb基因组序列草图。报道的分离物具有生产柔红素和β-胡萝卜素色素的潜力,并且含有一个假定的新型β-内酯的额外生物合成基因簇。基因组中还含有一个推测为β-内酰胺酶blaJOHN-1类似物的基因,并且存在一个推测为IS3家族的新插入序列的多个拷贝。该基因组增加了黄杆菌属的测序数据资源,可用于研究该属的微生物色素生产,抗微生物药物抗性基因和可移动遗传元件。
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引用次数: 0
Characterization of microbiome diversity unveils substantial microbial variation in mangrove soil sediments from coastal regions of Malaysia. 微生物组多样性的表征揭示了马来西亚沿海地区红树林土壤沉积物中大量微生物的变化。
Pub Date : 2025-06-18 eCollection Date: 2025-01-01 DOI: 10.1099/acmi.0.000902.v3
Prashantha Hebbar, Oh Bi Han, Ng Xin Yan, Dominic Kay, Kwa Yee Chu, James Sy-Keen Woon, Pang Kok Lun, Shama Prasada Kabekkodu, Alevoor S Bharath Prasad, Bharathi Prakash, Nadine Nograles, Mahibub Mahamadsa Kanakal, Michaela Goodson, Shubhada Nagaraja, Roshan Mascarenhas

The mangrove ecosystems are of great ecological importance found in tropical and subtropical coasts, including Malaysia. The microbial communities in the mangrove sediments play an indispensable role in maintaining homeostasis and supporting biodiversity. However, mangroves are facing various threats due to increasing anthropogenic activities. Thus, it is important to monitor the microbial community to improve our understanding of anthropogenic pressure on reshaping these ecosystems. This study examines the microbial community diversity in mangrove sediments of southern peninsular Malaysia. High-throughput MinION sequencing of the 16S rRNA gene was performed to compare the soil microbiome diversity in 35 samples from 8 different mangroves representing Sungai Sedili Kecil and Sungai Sedili Besar that flow into the South China Sea; Sungai Pulai, Sungai Melayu, Sungai Danga, Sungai Skudai and Sungai Johor that join the Straits of Johor; and Pulau Kukup from the Straits of Malacca. The metagenomic classification performed with 16S rRNA showed 2,573 taxa comprising 32 phyla. Total abundance analysis showed Pseudomonadota (67-69%), Bacteroidota (6-8%), Bacillota (5-8%), Campylobacterota (4-5%), Acidobacteriota (3-4%), Planctomycetota (2-4%) and Actinomycetota (1-2%) as the relatively common phyla. Alpha diversity indices revealed significantly higher richness in samples from mangroves of the South China Sea. Further, the 'Shannon' index showed a significant difference in diversity between Sungai Melayu and Sungai Pulai. Higher abundance of Burkholderiaceae, Bacillaceae and Enterobacteriaceae suggests a difference in the microbial community structure. This study stands as the first comprehensive analysis of microbial communities for future monitoring and conservation in these mangroves.

红树林生态系统在包括马来西亚在内的热带和亚热带海岸具有重要的生态意义。红树林沉积物中的微生物群落在维持生态平衡和支持生物多样性方面发挥着不可或缺的作用。然而,由于人类活动的增加,红树林正面临着各种威胁。因此,重要的是监测微生物群落,以提高我们对重塑这些生态系统的人为压力的理解。本研究考察了马来西亚半岛南部红树林沉积物中的微生物群落多样性。采用16S rRNA基因的高通量MinION测序方法,比较了流入南海的Sungai Sedili Kecil和Sungai Sedili Besar 8个不同红树林35个样品的土壤微生物群多样性;连接柔佛海峡的双溪蒲莱、双溪美莱、双溪丹加、双溪斯古岱、双溪柔佛;以及马六甲海峡的古姑岛。利用16S rRNA进行宏基因组分类,共发现32门2573个分类群。总丰度分析显示,假单胞菌门(67-69%)、拟杆菌门(6-8%)、杆状杆菌门(5-8%)、弯曲菌门(4-5%)、酸杆菌门(3-4%)、植物菌门(2-4%)和放线菌门(1-2%)是较为常见的门。α多样性指数显示南海红树林的丰富度显著高于其他红树林。此外,“香农”指数显示双盖马来岛和双盖蒲莱岛的多样性存在显著差异。burkholderaceae、Bacillaceae和Enterobacteriaceae丰度较高,表明微生物群落结构存在差异。这项研究是第一次对这些红树林微生物群落进行全面分析,为未来的监测和保护提供依据。
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引用次数: 0
Effect of laboratory manual layout: does experiential learning benefit from authentic context? 实验室手册布局的效果:体验式学习是否受益于真实情境?
Pub Date : 2025-06-17 eCollection Date: 2025-01-01 DOI: 10.1099/acmi.0.000955.v4
Victoria Alice Kate Easton

Experiential learning is the pedagogic foundation of practical laboratory education. This process of learning through experience enables students to develop a deeper understanding of the theoretical material as well as valuable real-world skills. However, there is often a disconnect between the authentic, real-world context of performing laboratory skills and the method of instruction within higher education. This study developed two student laboratory manuals; one which followed a traditionally linear 'week by week' format, and another which took inspiration from a publication format and listed the protocols in a distinct 'methods' section. The effect the change of layout had on student learning was assessed through analysis of student summative assessment and interaction with the online learning environment. Additionally, the effect on student confidence and perceived technical skills development was assessed through a student survey. The differences in layout resulted in no significant differences in student assessment performance but did result in higher levels of engagement with the online learning environment. The student survey reported an increase in technical confidence (21%) and skill (31%) with the authentic 'methods' section layout changes compared to the traditional format. The increase in student engagement, confidence and perceived skill shows that experiential learning benefits from placing the information in an authentic context.

体验式学习是实践实验教育的教学基础。这种通过经验学习的过程使学生对理论材料有了更深的理解,同时也获得了宝贵的现实技能。然而,在执行实验室技能的真实世界背景与高等教育中的教学方法之间往往存在脱节。这项研究开发了两本学生实验室手册;一个遵循传统的线性“周接周”格式,另一个从出版物格式中获得灵感,并在不同的“方法”部分列出了协议。通过对学生总结性评价的分析和与在线学习环境的互动来评估布局变化对学生学习的影响。此外,通过学生调查评估了对学生信心和感知技术技能发展的影响。布局的差异并没有导致学生在评估表现上的显著差异,但确实导致了学生对在线学习环境的更高参与度。学生调查报告说,与传统格式相比,真实的“方法”部分布局的变化增加了技术信心(21%)和技能(31%)。学生参与度、信心和感知技能的提高表明,体验式学习受益于将信息置于真实的环境中。
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引用次数: 0
Culture harder: use more specimens to increase methicillin-resistant Staphylococcus aureus culture yield relative to PCR. 培养难度:相对于PCR,使用更多的标本来增加耐甲氧西林金黄色葡萄球菌的培养产量。
Pub Date : 2025-06-03 eCollection Date: 2025-01-01 DOI: 10.1099/acmi.0.000918.v4
Arvette E Mitchell, Arpit P Patel, Jennifer DiCandilo, Zachary W Rebollido, Matthew A Pettengill

Methicillin-resistant Staphylococcus aureus (MRSA) causes considerable morbidity and mortality in both community-acquired and healthcare-associated infections, but detecting colonization with MRSA has been shown to improve patient outcomes in certain clinical settings. MRSA colonization detection has been carried out in a variety of ways, with molecular assays having superior sensitivity in most studies relative to culture, but culture is disadvantaged in some comparisons by utilization of low specimen volumes. We compared a commercial molecular assay to both low-volume (10 µl) and high-volume (650 µl) cultures and found that increasing the volume utilized for culture led to the detection of 25% more cases than low-volume culture.

耐甲氧西林金黄色葡萄球菌(MRSA)在社区获得性感染和医疗保健相关感染中引起相当大的发病率和死亡率,但在某些临床环境中,检测MRSA定植已被证明可以改善患者的预后。MRSA定植检测的方法多种多样,相对于培养,分子测定法在大多数研究中具有更高的灵敏度,但培养法在某些比较中由于利用低标本体积而处于劣势。我们将商业分子检测与小体积(10µl)和大体积(650µl)培养进行了比较,发现增加用于培养的体积比小体积培养多检测25%的病例。
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引用次数: 0
A cautionary tale of false-positive histoplasma urine antigen in an HIV patient: a case report. 一个HIV患者尿组织浆抗原假阳性的警示故事:一个病例报告。
Pub Date : 2025-05-28 eCollection Date: 2025-01-01 DOI: 10.1099/acmi.0.000929.v3
Mohammad Z Khrais, Jake Smith, Tanmay Gandhi, Shahrukh Arif, Juan Carlos Rico

Introduction. Coccidioidomycosis, or Valley fever, is a fungal disease caused by Coccidioides species, prevalent in parts of the southwestern United States. It usually results from inhaling spores from soil and is a common cause of pneumonia in these regions. Case Presentation. We present a unique case of coccidioidomycosis in an immunodeficient male patient secondary to human immunodeficiency virus infection with poor adherence to anti-retroviral treatment. After presenting with non-specific symptoms and pre-syncope, he was initially diagnosed with pneumonia based on chest X-ray findings, but his symptoms failed to improve with antibiotics. He was treated for presumed pulmonary histoplasmosis following a positive histoplasma urine antigen test. However, the patient worsened clinically. Following a computed tomography scan demonstrating a large necrotic lung consolidation, fungal stain and culture of tissue biopsied through endobronchial ultrasound confirmed coccidioidomycosis. The patient received 2 weeks of liposomal amphotericin with clinical improvement before discharge with itraconazole. Conclusion. The histoplasma antigen test can be falsely positive due to cross-reaction with other fungal infections like blastomycosis, paracoccidioidomycosis or talaromycosis, and less frequently, coccidioidomycosis or aspergillosis. Diagnosis of coccidioidomycosis requires a high index of suspicion outside the expected geographic distribution in the appropriate clinical setting. Our case highlights the risk of false-positive antigen test results and the importance of invasive diagnostics, including bronchoscopy to obtain fungal cultures, if the diagnosis remains uncertain.

介绍。球孢子菌病,或谷热,是一种由球孢子菌引起的真菌疾病,在美国西南部部分地区流行。它通常是由吸入土壤中的孢子引起的,是这些地区肺炎的常见原因。例演示。我们提出了一个独特的球孢子菌病的免疫缺陷男性患者继发于人类免疫缺陷病毒感染与抗逆转录病毒治疗依从性差。在出现非特异性症状和晕厥前期后,根据胸部x光检查结果,他最初被诊断为肺炎,但他的症状在抗生素治疗后未能改善。在尿组织浆体抗原检测呈阳性后,他被诊断为肺组织浆菌病。然而,患者在临床上病情恶化。计算机断层扫描显示肺大坏死实变,真菌染色和支气管超声活检组织培养证实球孢子菌病。患者接受两性霉素脂质体治疗2周,临床情况好转,出院前用伊曲康唑治疗。结论。由于与其它真菌感染,如芽孢菌病、副球孢子菌病或talaromycosis,以及较少出现的球孢子菌病或曲霉病的交叉反应,组织浆抗原试验可能出现假阳性。球孢子菌病的诊断需要在适当的临床环境中对预期地理分布之外的高度怀疑。我们的病例强调了假阳性抗原检测结果的风险和侵入性诊断的重要性,包括支气管镜检查获得真菌培养,如果诊断仍然不确定。
{"title":"A cautionary tale of false-positive histoplasma urine antigen in an HIV patient: a case report.","authors":"Mohammad Z Khrais, Jake Smith, Tanmay Gandhi, Shahrukh Arif, Juan Carlos Rico","doi":"10.1099/acmi.0.000929.v3","DOIUrl":"10.1099/acmi.0.000929.v3","url":null,"abstract":"<p><p><b>Introduction.</b> Coccidioidomycosis, or Valley fever, is a fungal disease caused by <i>Coccidioides</i> species, prevalent in parts of the southwestern United States. It usually results from inhaling spores from soil and is a common cause of pneumonia in these regions. <b>Case Presentation.</b> We present a unique case of coccidioidomycosis in an immunodeficient male patient secondary to human immunodeficiency virus infection with poor adherence to anti-retroviral treatment. After presenting with non-specific symptoms and pre-syncope, he was initially diagnosed with pneumonia based on chest X-ray findings, but his symptoms failed to improve with antibiotics. He was treated for presumed pulmonary histoplasmosis following a positive histoplasma urine antigen test. However, the patient worsened clinically. Following a computed tomography scan demonstrating a large necrotic lung consolidation, fungal stain and culture of tissue biopsied through endobronchial ultrasound confirmed coccidioidomycosis. The patient received 2 weeks of liposomal amphotericin with clinical improvement before discharge with itraconazole. <b>Conclusion.</b> The histoplasma antigen test can be falsely positive due to cross-reaction with other fungal infections like blastomycosis, paracoccidioidomycosis or talaromycosis, and less frequently, coccidioidomycosis or aspergillosis. Diagnosis of coccidioidomycosis requires a high index of suspicion outside the expected geographic distribution in the appropriate clinical setting. Our case highlights the risk of false-positive antigen test results and the importance of invasive diagnostics, including bronchoscopy to obtain fungal cultures, if the diagnosis remains uncertain.</p>","PeriodicalId":94366,"journal":{"name":"Access microbiology","volume":"7 5","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12120142/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144182686","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}
引用次数: 0
Are reads required? High-precision variant calling from bacterial genome assemblies. 是否需要阅读?来自细菌基因组组装的高精度变体调用。
Pub Date : 2025-05-28 eCollection Date: 2025-01-01 DOI: 10.1099/acmi.0.001025.v3
Ryan R Wick, Louise M Judd, Timothy P Stinear, Ian R Monk

Accurate nucleotide variant calling is essential in microbial genomics, particularly for outbreak tracking and phylogenetics. This study evaluates variant calls derived from genome assemblies compared to traditional read-based variant-calling methods, using seven closely related Staphylococcus aureus isolates sequenced on Illumina and Oxford Nanopore Technologies platforms. By benchmarking multiple assembly and variant-calling pipelines against a ground truth dataset, we found that read-based methods consistently achieved high accuracy. Assembly-based approaches performed well in some cases but were highly dependent on assembly quality, as errors in the assembly led to false-positive variant calls. These findings underscore the need for improved assembly techniques before the potential benefits of assembly-based variant calling (such as reduced computational requirements and simpler data management) can be realized.

准确的核苷酸变异识别在微生物基因组学中是必不可少的,特别是在疫情跟踪和系统发育中。本研究利用在Illumina和Oxford Nanopore Technologies平台上测序的7株密切相关的金黄色葡萄球菌分离株,与传统的基于读取的变异召唤方法相比,评估了来自基因组组装的变异召唤。通过对多个装配和变量调用管道进行基准测试,我们发现基于读取的方法始终具有较高的准确性。基于程序集的方法在某些情况下表现良好,但高度依赖于程序集质量,因为程序集中的错误会导致误报变量调用。这些发现强调,在实现基于程序集的变量调用的潜在好处(例如减少计算需求和简化数据管理)之前,需要改进装配技术。
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引用次数: 0
Crystalline lens dislocation as a presenting sign of Streptococcus pyogenes invasive infections. 晶状体脱位是化脓性链球菌侵袭性感染的表现。
Pub Date : 2025-05-27 eCollection Date: 2025-01-01 DOI: 10.1099/acmi.0.000903.v4
Paulina Liberman, Jacob G Light, Shazia Dharssi, Tracy Howard, Jennifer Lu, Patricia J Simner, Karen C Carroll, Cheng-Ying Ho

Introduction. To describe two cases of crystalline lens dislocation as a presenting feature of invasive group A Streptococcus (GAS) infection and its management. Case presentation. We report on a 58-year-old woman and a 36-year-old man who presented in 2024 with acute vision loss and severe ocular and systemic symptoms. Both patients were found to have lens dislocation and were diagnosed with invasive GAS infection. The 58-year-old woman had a complicated clinical course leading to enucleation, while the 36-year-old man responded favourably to early and aggressive treatment with systemic and intravitreal antibiotics. The responsible GAS strains were sequence type (ST) 28 and ST433, respectively. Conclusion. These cases highlight the importance of recognizing crystalline lens dislocation as a potential sign of ocular GAS infection. Two specific strain types of GAS associated with these findings, ST28 and ST433, are reported. In patients with GAS sepsis presenting with corneal oedema and zonular loss, clinicians should immediately initiate treatment, including intravitreal antibiotic injections and systemic therapy. Prompt and aggressive management can be crucial in preserving ocular structures.

介绍。描述两例浸润性a群链球菌(GAS)感染的晶状体脱位及其治疗。例演示。我们报告了一名58岁的女性和一名36岁的男性,他们在2024年出现了急性视力丧失和严重的眼部和全身症状。两例患者均发现晶状体脱位,并被诊断为浸润性GAS感染。这名58岁的女性有一个复杂的临床过程导致了眼球摘除,而这名36岁的男性对早期和积极的全身和玻璃体内抗生素治疗反应良好。负责的GAS菌株分别为序列型(ST) 28和ST433。结论。这些病例强调了识别晶状体脱位作为眼部GAS感染的潜在征兆的重要性。报道了与这些发现相关的两种特定的GAS菌株类型,ST28和ST433。对于以角膜水肿和角膜带丢失为表现的GAS脓毒症患者,临床医生应立即开始治疗,包括玻璃体内抗生素注射和全身治疗。及时和积极的治疗对于保护眼部结构至关重要。
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引用次数: 0
Multi-metal-resistant Staphylococcus warneri strain TWSL_1: revealing heavy metal-resistant genomic features by whole-genome sequencing and analysis. 耐多金属瓦纳利葡萄球菌菌株TWSL_1:通过全基因组测序和分析揭示耐重金属基因组特征
Pub Date : 2025-05-27 eCollection Date: 2025-01-01 DOI: 10.1099/acmi.0.000954.v5
Dilani Chathurika Dissanayake, Naduviladath Vishvanath Chandrasekharan, Champika Dilrukshi Wijayarathna

This study explores the genomic basis of heavy metal resistance in Staphylococcus warneri strain TWSL_1, isolated from industrial textile effluent. The strain exhibited strong resistance to Cd²+, Pb²+ and Cu²+, with MICs of 50, 1,200 and 75 mg l-1, respectively. Whole-genome sequencing revealed a 2.66 Mb genome with 2,567 CDSs and a 99.81% average nucleotide identity to S. warneri WS479. Comparative genomic analysis at the genus level revealed that S. warneri strain TWSL_1 possesses a unique and expanded repertoire of heavy metal resistance genes, including the cadmium efflux system accessory protein and cadmium resistance protein, which are absent in pathogenic Staphylococcus sp. used for the comparison. Phylogenetic analysis confirmed its classification within S. warneri, with strong bootstrap support (100). Functional annotation highlighted metabolic versatility and stress response capabilities, supporting its adaptation to metal-rich environments. S. warneri TWSL_1 demonstrated high Pb²+ removal efficiency, reducing concentrations by over 70%. These findings highlight S. warneri TWSL_1 as a promising candidate for heavy metal bioremediation with potential applications in environmental detoxification and monitoring strategies.

本研究探讨了从工业纺织废水中分离的瓦纳里葡萄球菌TWSL_1对重金属抗性的基因组基础。菌株对Cd²+、Pb²+和Cu²+具有较强的抗性,mic分别为50、1200和75 mg l-1。全基因组测序结果显示,该基因组全长2.66 Mb,包含2,567个cds,平均核苷酸与S. warneri WS479同源性为99.81%。在属水平上的比较基因组分析表明,warneri菌株TWSL_1具有独特和扩展的重金属抗性基因库,包括镉外排系统辅助蛋白和镉抗性蛋白,这是用于比较的致病性葡萄球菌所缺乏的。系统发育分析证实其属于S. warneri,具有很强的bootstrap支持(100)。功能注释强调了代谢的多功能性和应激反应能力,支持其适应富金属环境。S. warneri TWSL_1表现出较高的Pb²+去除率,浓度降低70%以上。这些发现突出了S. warneri TWSL_1作为重金属生物修复的潜在候选者,在环境解毒和监测策略方面具有潜在的应用前景。
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引用次数: 0
Derivatization of pBACpAK entrapment vectors for enhanced mobile genetic element transposition detection in multidrug-resistant Escherichia coli. 多药耐药大肠杆菌移动基因转位检测pBACpAK包埋载体的衍生化。
Pub Date : 2025-05-23 eCollection Date: 2025-01-01 DOI: 10.1099/acmi.0.001013.v3
Supathep Tansirichaiya, Wasawat Leartsiwawinyu, Nattharee Thanawan, Richard N Goodman, Chanwit Tribuddharat, Adam P Roberts

Aim. Antimicrobial resistance poses a critical global health threat, driven by the dissemination of resistance genes via mobile genetic elements (MGEs). This study aims to enhance the detection of MGE insertions in multidrug-resistant Escherichia coli by derivatizing the pBACpAK entrapment vector. Methods and results. Three derivatives were constructed with additional nucleotides upstream of the cI repressor gene, based on conserved regions identified from GenBank sequences containing known IS26 and IS1 insertions. Using colony PCR, intracellular transposition screening was performed on 194 tetracycline-resistant colonies from four E. coli ESI123 strains carrying different pBACpAK constructs. The derivatives showed increased MGE capture rates (10.7-73.1 %) compared to the WT vector (3.75%), identifying multiple MGEs, including the novel composite transposon Tn7824. Tn7824 harbours the bla OXA-181 carbapenem resistance gene and the qnrS1 quinolone resistance gene, highlighting the clinical relevance of these findings. Long-read sequencing of transposants confirmed the accuracy of MGE identification and structural characterization, which also revealed chromosomal integration events of the pBACpAK derivatives mediated by flanking insertion sequences. Conclusions. The modifications introduced in the pBACpAK derivatives could increase the detection of transposition events by alleviating spatial constraints, allowing for more robust MGE detection.

的目标。由于耐药性基因通过移动遗传元件(MGEs)传播,抗菌素耐药性对全球健康构成严重威胁。本研究旨在通过衍生化pBACpAK包埋载体,提高MGE插入在耐多药大肠杆菌中的检测能力。方法与结果。基于从GenBank序列中鉴定出的含有已知IS26和IS1插入的保守区域,在cI抑制基因上游添加了额外的核苷酸,构建了三个衍生物。采用集落PCR技术,对携带不同pBACpAK构建体的4株大肠杆菌ESI123的194个四环素耐药菌落进行了细胞内转位筛选。与WT载体(3.75%)相比,该衍生物显示出更高的MGE捕获率(10.7% - 73.1%),识别出多个MGE,包括新型复合转座子Tn7824。Tn7824含有bla OXA-181碳青霉烯类耐药基因和qnrS1喹诺酮类耐药基因,突出了这些发现的临床相关性。转座的长读测序证实了MGE鉴定和结构表征的准确性,这也揭示了pBACpAK衍生物由侧翼插入序列介导的染色体整合事件。结论。在pBACpAK衍生物中引入的修改可以通过减轻空间限制来增加对转位事件的检测,从而实现更稳健的MGE检测。
{"title":"Derivatization of pBACpAK entrapment vectors for enhanced mobile genetic element transposition detection in multidrug-resistant Escherichia coli.","authors":"Supathep Tansirichaiya, Wasawat Leartsiwawinyu, Nattharee Thanawan, Richard N Goodman, Chanwit Tribuddharat, Adam P Roberts","doi":"10.1099/acmi.0.001013.v3","DOIUrl":"10.1099/acmi.0.001013.v3","url":null,"abstract":"<p><p><b>Aim.</b> Antimicrobial resistance poses a critical global health threat, driven by the dissemination of resistance genes via mobile genetic elements (MGEs). This study aims to enhance the detection of MGE insertions in multidrug-resistant <i>Escherichia coli</i> by derivatizing the pBACpAK entrapment vector. <b>Methods and results.</b> Three derivatives were constructed with additional nucleotides upstream of the <i>cI</i> repressor gene, based on conserved regions identified from GenBank sequences containing known IS<i>26</i> and IS<i>1</i> insertions. Using colony PCR, intracellular transposition screening was performed on 194 tetracycline-resistant colonies from four <i>E. coli</i> ESI123 strains carrying different pBACpAK constructs. The derivatives showed increased MGE capture rates (10.7-73.1 %) compared to the WT vector (3.75%), identifying multiple MGEs, including the novel composite transposon Tn<i>7824</i>. Tn<i>7824</i> harbours the <i>bla</i> <sub>OXA-181</sub> carbapenem resistance gene and the <i>qnrS1</i> quinolone resistance gene, highlighting the clinical relevance of these findings. Long-read sequencing of transposants confirmed the accuracy of MGE identification and structural characterization, which also revealed chromosomal integration events of the pBACpAK derivatives mediated by flanking insertion sequences. <b>Conclusions.</b> The modifications introduced in the pBACpAK derivatives could increase the detection of transposition events by alleviating spatial constraints, allowing for more robust MGE detection.</p>","PeriodicalId":94366,"journal":{"name":"Access microbiology","volume":"7 5","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12102499/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144145347","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}
引用次数: 0
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