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“Reflexions on the role, diversity, conservation and management of the genetic microbial resources in Agriculture” 关于农业遗传微生物资源的作用、多样性、保护和管理的思考
IF 4.8 Q1 MICROBIOLOGY Pub Date : 2025-01-01 DOI: 10.1016/j.crmicr.2025.100365
María del Pilar Rodríguez Guzmán , Ismael Fernando Chávez Díaz , Lily Xochilt Zelaya Molina
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引用次数: 0
Environmental biocontamination by SARS-CoV-2 Virus in the hospital setting SARS-CoV-2病毒对医院环境的生物污染
IF 4.8 Q1 MICROBIOLOGY Pub Date : 2025-01-01 DOI: 10.1016/j.crmicr.2025.100355
M Espejo Mambié , D San Jose-Saras , C Bischofberger Valdés , C Díaz-Agero Pérez , JC Galán Montemayor , L Martínez-García , M Abreu Di-Berardino , P Moreno-Nunez , J Vicente-Guijarro , J.M Aranaz-Andrés

Background

Demonstrating the capability to isolate biological material from the environment was fundamental to supporting any transmission route. Various and inconsistent methodologies have been used to address this issue; however, the debate in scientific societies about the possibility of airborne transmission as a source of SARS-CoV-2 spread remained open.

Objective

To analyze SARS-CoV-2 contamination in the air and on surfaces in a hospital setting during the COVID-19 pandemic.

Methods

This study involved air and surface sampling in the emergency, hospitalization, and intensive care unit areas of the Ramón y Cajal University Hospital. A consistent methodology was used for all samples, and clinical and environmental parameters and characterization of each location were recorded.

Results

A total of 234 samples were collected, comprising 160 surface samples and 74 air samples, of which 6.84 % tested positive (13/160 surface samples and 3/74 air samples). High-contact surfaces had the highest proportion of positive samples (12/13). All positive air samples were identified within 2 m of patients who had recently developed symptoms (<5 days). High dependency and elevated temperatures seemed to indicate a higher risk of environmental biocontamination. Additionally, there was a higher risk of contamination in the intensive care units than in the hospitalization or emergency units.
证明从环境中分离生物材料的能力是支持任何传播途径的基础。已经使用了各种不一致的方法来解决这个问题;然而,科学界关于空气传播作为SARS-CoV-2传播来源的可能性的争论仍然存在。目的分析2019冠状病毒病大流行期间医院空气和表面的SARS-CoV-2污染情况。方法本研究采用Ramón y Cajal大学医院急诊、住院和重症监护病房区域的空气和地面采样。对所有样本采用一致的方法,记录临床和环境参数以及每个位置的特征。结果共采集标本234份,其中地表标本160份,空气标本74份,阳性率6.84%(地表标本13/160,空气标本3/74)。高接触面阳性样品比例最高(12/13)。所有阳性空气样本均在最近出现症状(5天)的患者2米范围内确定。高度依赖和高温似乎表明环境生物污染的风险更高。此外,重症监护病房的污染风险高于住院或急诊病房。
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引用次数: 0
Multiple influences on cyanobacterial abundance and diversity in the Beijing-Tianjin-Hebei Economic Circle and nearby areas of China 京津冀经济圈及周边地区蓝藻丰度和多样性的多重影响
IF 4.8 Q1 MICROBIOLOGY Pub Date : 2025-01-01 DOI: 10.1016/j.crmicr.2025.100400
Wei Wang, Hanjie Huang, Kangxu Zhao, Junping Lv, Xudong Liu, Shulian Xie, Jia Feng
In recent years, there has been a marked increase in the frequency and duration of cyanobacterial blooms, which has attracted widespread attention from all sectors of society. However, the processes and pathways leading to the formation of cyanobacterial blooms are largely stochastic, and a targeted quantitative model incorporating multiple drivers is required. A series of fieldwork and analytical procedures were conducted on 11 lakes in Beijing-Tianjin-Hebei and neighboring regions. The objective of this research was to investigate the effects of location, climate, habitat, metabolism and biology on the abundance of cyanobacteria. The findings indicated that location factors exerted an indirect influence on cyanobacterial density, with a calculated effect size of −0.001. This observation is consistent with the documented prevalence of cyanobacterial bloom outbreaks, suggesting that cyanobacterial abundance is not directly associated with location. Furthermore, the water environment (0.35 indirect and −0.37 direct effects) and climatic conditions (−0.2 indirect and 0.16 direct effects) had significant direct and indirect effects on cyanobacterial cell densities. It was also determined that high nutrients and high temperatures remained the basis for promoting cyanobacterial blooms. It is noteworthy that the microbial community exerted a substantial inhibitory effect on the proliferation of the cyanobacterial community (1.13 direct inhibitory effect). It is hypothesized that the negative effect is derived from two primary sources: the competition for ecological niches by organisms exhibiting similarity, and the direct inhibitory effect of certain bacteria on the growth of cyanobacteria, a negative correlation between acidobacteriota, bacteroide and a variety of cyanobacteria was observed in our study. The effect of specific bacteria is found to be significantly negatively correlated with the prevalence of a wide range of cyanobacteria, which is consistent with the results of many extant studies. These extant studies indicate that the formation of cyanobacterial communities is primarily influenced by microorganisms in the environment. The results of this study will provide new and complementary evidence on the mechanisms of cyanobacterial bloom occurrence and development and the design of management strategies.
近年来,蓝藻爆发的频率和持续时间明显增加,引起了社会各界的广泛关注。然而,导致蓝藻华形成的过程和途径在很大程度上是随机的,需要一个包含多个驱动因素的有针对性的定量模型。对京津冀及周边地区的11个湖泊进行了一系列野外调查和分析。本研究的目的是调查地点、气候、栖息地、新陈代谢和生物学对蓝藻丰度的影响。研究结果表明,位置因素对蓝藻密度有间接影响,计算的效应值为−0.001。这一观察结果与记录的蓝藻爆发流行一致,表明蓝藻丰度与位置没有直接关系。水环境(间接效应为0.35,直接效应为- 0.37)和气候条件(间接效应为- 0.2,直接效应为0.16)对蓝藻细胞密度有显著的直接和间接影响。研究还确定,高营养和高温仍然是促进蓝藻繁殖的基础。值得注意的是,微生物群落对蓝藻群落的增殖具有实质性的抑制作用(1.13直接抑制作用)。假设这种负面影响主要来自两个方面:具有相似性的生物对生态位的竞争,以及某些细菌对蓝藻生长的直接抑制作用,在我们的研究中观察到酸杆菌群、拟杆菌群与多种蓝藻之间存在负相关。研究发现,特定细菌的作用与多种蓝藻的流行率呈显著负相关,这与许多现有研究的结果一致。这些现存的研究表明,蓝藻群落的形成主要受到环境中微生物的影响。本研究结果将为蓝藻华的发生发展机制和管理策略的设计提供新的补充证据。
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引用次数: 0
Screening and discovery of an antiviral candidate inhibiting the SARS-CoV-2 envelope (2-E) channel 抑制SARS-CoV-2包膜(2-E)通道的抗病毒候选药物的筛选和发现
IF 4.8 Q1 MICROBIOLOGY Pub Date : 2025-01-01 DOI: 10.1016/j.crmicr.2025.100409
Han Zhang , Shuxin Shi , Lujia Sun , Shuangqu Li , Yan Zhang , Ziyue Li , Jingjing Hou , Pingan Li , Jingshan Shen , Xi Cheng , Shibo Jiang , Zhaobing Gao , Xinling Wang , Xiangrui Jiang , Bingqing Xia
The SARS-CoV-2-encoded 2-E channel is critical in the viral life cycle and pathogenesis. By facilitating viral replication, it promotes the dysregulation of inflammatory pathways, leading to cytokine storm, and triggers DNA damage response (DDR), thus exacerbating disease progression. The 2-E channel, a viroporin, is a promising antiviral target. However, the lack of specific inhibitors and effective screening methods has hindered therapeutic exploitation of the 2-E channel. To address this gap, we report on a fluorescence-based screening assay that targets the 2-E channel activity, resulting in the identification of potential inhibitory molecules. After performing both electrophysiological studies and surface plasmon resonance (SPR) analyses, we identified the top-ranked candidate, TPN10518, as a pore-blocking inhibitor of the 2-E channel. TPN10518 binds to a hydrophobic pocket in the C-terminal vestibule of the 2-E channel, thereby inhibiting its activity. Functional evaluation showed that TPN10518 exhibits significant antiviral efficacy in vitro, while, at the same time, effectively protecting against 2-E channel-mediated host damage and suppressing cytokine storm caused by dysregulated homeostasis of inflammatory pathways in vivo. Therefore, our work introduces a screening method for targeting 2-E channels, establishes the 2-E channel as a viable therapeutic target against SARS-CoV-2, and identifies TPN10518 as a promising antiviral candidate.
sars - cov -2编码的2-E通道在病毒生命周期和发病机制中至关重要。通过促进病毒复制,它促进炎症通路失调,导致细胞因子风暴,触发DNA损伤反应(DDR),从而加剧疾病进展。2-E通道,一种病毒蛋白,是一种很有前途的抗病毒靶点。然而,缺乏特异性抑制剂和有效的筛选方法阻碍了2-E通道的治疗利用。为了解决这一差距,我们报告了一种基于荧光的筛选试验,该试验针对2-E通道活性,从而鉴定出潜在的抑制分子。在进行了电生理研究和表面等离子体共振(SPR)分析后,我们确定了排名第一的候选药物TPN10518,作为2-E通道的孔阻断抑制剂。TPN10518结合到2-E通道c端前庭的疏水口袋上,从而抑制其活性。功能评价表明,TPN10518在体外具有显著的抗病毒作用,同时在体内可有效保护2-E通道介导的宿主损伤,抑制炎症通路稳态失调引起的细胞因子风暴。因此,我们的工作引入了一种靶向2-E通道的筛选方法,确定了2-E通道是针对SARS-CoV-2的可行治疗靶点,并确定了TPN10518是一种有希望的抗病毒候选药物。
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引用次数: 0
Cell wall polysaccharide alters the surface electric potential of Lacticaseibacillus paracasei strain Shirota and enhances acid tolerance 细胞壁多糖改变副干酪乳杆菌Shirota菌株的表面电位,增强其耐酸能力
IF 4.8 Q1 MICROBIOLOGY Pub Date : 2025-01-01 DOI: 10.1016/j.crmicr.2025.100430
Kosuke Kato, Madoka Nakamura, Masaki Serata, Takekazu Okumura
LCPS-1 is a cell wall polysaccharide (CWPS) of Lacticaseibacillus paracasei (formerly Lactobacillus casei) strain Shirota (LcS). LCPS-1 enhances the acid tolerance of LcS; however, the mechanism of action remains unclear. This study aimed to elucidate how LCPS-1 enhances LcS growth and survivability under acidic conditions. When cultured in media acidified with hydrochloric, lactic, or acetic acid, wild-type LcS showed significantly more growth than an LCPS-1-deficient mutant strain (p < 0.01), indicating that LCPS-1 confers enhanced protection against proton (H+) stress. Wild-type LcS also exhibited a significant 73–83 % reduction (p < 0.01) in negative surface potential compared to that of the mutant. Additionally, wild-type LcS showed significantly more growth than the mutant (p < 0.01) in the presence of cupric oxide, which releases bactericidal Cu2+ ions. In contrast, wild-type LcS was slightly more susceptible than the mutant to lethal concentrations of hydroxide ions. These findings suggest that LCPS-1 modulates the surface electric potential of LcS cells, thereby influencing the interaction strength between the cell walls and environmental ions, ultimately altering the tolerance of the cells to ionic stress factors. These findings provide crucial insights into the role of CWPSs in modulating the surface electric potential of microorganisms, which affects microbial interactions with environmental ions. This enhanced understanding of CWPS function enables the development of more resilient probiotic strains, potentially improving their efficacy in various industrial and therapeutic applications.
LCPS-1是副干酪乳杆菌(Lactobacillus paracasei,原干酪乳杆菌)Shirota菌株的细胞壁多糖(CWPS)。LCPS-1增强LcS的耐酸能力;然而,其作用机制尚不清楚。本研究旨在阐明LCPS-1如何在酸性条件下促进LcS的生长和存活。当在盐酸、乳酸或醋酸酸化的培养基中培养时,野生型LcS的生长速度明显高于lcps -1缺陷突变株(p <;0.01),表明LCPS-1对质子(H+)胁迫具有增强的保护作用。野生型LcS也表现出73 - 83%的显著减少(p <;与突变体相比,负表面电位差异为0.01)。此外,野生型LcS的生长显著高于突变型(p <;0.01)在氧化铜存在下,释放出杀菌Cu2+离子。相比之下,野生型LcS对氢氧离子致死浓度的敏感性略高于突变型LcS。这些发现表明,LCPS-1调节LcS细胞的表面电位,从而影响细胞壁与环境离子之间的相互作用强度,最终改变细胞对离子应激因子的耐受性。这些发现为cwps在调节微生物表面电位中的作用提供了重要的见解,而微生物表面电位影响微生物与环境离子的相互作用。对CWPS功能的深入了解有助于开发更具弹性的益生菌菌株,潜在地提高其在各种工业和治疗应用中的功效。
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引用次数: 0
The Jarman–Bell principle revisited: Gut microbiota facilitate body size-dependent nutritional strategies in herbivores 重新审视贾曼-贝尔原理:肠道微生物群促进了草食动物体型依赖的营养策略
IF 5.8 Q1 MICROBIOLOGY Pub Date : 2025-01-01 DOI: 10.1016/j.crmicr.2025.100508
Peng Zheng , Peng Ding , Wei-Zhen Gao , Xi Chen , Lei Shi
Body size is a key determinant of nutritional strategies in herbivores, yet the role of gut microbiota in mediating these strategies remains insufficiently explored. To address this gap, we examined two sympatric ungulates of contrasting body sizes in an arid ecosystem—red deer (Cervus elaphus, large-bodied) and goitered gazelle (Gazella subgutturosa, small-bodied)—to test three predictions derived from the Jarman–Bell principle (JBP). We applied dietary DNA metabarcoding, plant nutritional profiling, and 16S rRNA sequencing of gut microbiota to assess how body size shapes macronutrient intake, microbial composition, and functional adaptations. Both species exhibited similar macronutrient ratios dominated by carbohydrates (∼88–90 %), supporting the nutritional balancing hypothesis. However, they differed in nutrient targets and microbial profiles: red deer consumed more non-structural carbohydrates and fats, with enriched gut microbial taxa (Paludibacter, Turicibacter) linked to energy metabolism, reflecting an energy maximization strategy. In contrast, goitered gazelles ingested more crude protein and harbored microbial taxa (Prevotella) associated with protein digestion, alongside immune-supporting microbes (Butyricicoccus, Coprococcus, and Victivallis), consistent with a protein maximization strategy. Red deer also demonstrated a greater microbial capacity for fiber degradation (Fibrobacter, Ruminococcus albus) and detoxification of plant secondary metabolites (Clostridium, Desulfovibrio, Prevotella, Variovorax). Functional pathway analysis revealed enrichment in lignocellulose and terpenoid metabolism, indicating an adaptation to low-quality forage. In contrast, goitered gazelles showed limited microbial associations with fiber or phytotoxin regulation, suggesting weaker microbial-mediated adaptation to low-quality diets. These results demonstrate that body size governs distinct nutritional strategies in sympatric herbivores, mediated through differential gut microbiota composition and function. Larger species, such as red deer, exhibit enhanced microbial capacity for fiber and toxin regulation, allowing them to utilize lower-quality forage more efficiently.
体型是食草动物营养策略的关键决定因素,但肠道微生物群在调节这些策略中的作用仍未得到充分探讨。为了解决这一差距,我们研究了干旱生态系统中两种身体大小不同的同域有蹄类动物——马鹿(Cervus elaphus,大型)和喉羚(Gazella subgutturrosa,小型)——以验证从贾曼-贝尔原理(JBP)中得出的三个预测。我们应用膳食DNA元条形码、植物营养谱和肠道微生物群16S rRNA测序来评估体型如何影响宏量营养素摄入、微生物组成和功能适应。这两个物种都表现出相似的以碳水化合物为主的常量营养素比例(~ 88 - 90%),支持营养平衡假说。然而,它们在营养目标和微生物特征上存在差异:马鹿消耗更多的非结构性碳水化合物和脂肪,与能量代谢相关的肠道微生物类群(Paludibacter, Turicibacter)丰富,反映了能量最大化策略。相比之下,甲状腺瞪羚摄入了更多的粗蛋白质,并拥有与蛋白质消化相关的微生物类群(普雷沃氏菌),以及支持免疫的微生物(丁酸球菌、粪球菌和维氏菌),这与蛋白质最大化策略一致。马鹿在纤维降解(纤维杆菌、白色瘤胃球菌)和植物次生代谢物(梭状芽孢杆菌、脱硫弧菌、普雷沃氏菌、变异弧菌)解毒方面也表现出更强的微生物能力。功能途径分析显示其木质纤维素和萜类代谢富集,表明其适应低品质饲料。相比之下,甲状腺瞪羚与纤维或植物毒素调节的微生物关联有限,表明微生物介导的对低质量饮食的适应较弱。这些结果表明,同域食草动物的身体大小通过不同的肠道微生物群组成和功能来调节不同的营养策略。较大的物种,如马鹿,表现出增强的纤维和毒素调节的微生物能力,使它们能够更有效地利用低质量的饲料。
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引用次数: 0
Harnessing Lactobacillus-derived SCFAs for food and health: Pathways, genes, and functional implications 利用乳酸杆菌衍生的SCFAs用于食品和健康:途径、基因和功能意义
IF 5.8 Q1 MICROBIOLOGY Pub Date : 2025-01-01 DOI: 10.1016/j.crmicr.2025.100496
Yousef Nami , Milad Shaghaghi Ranjbar , Mahmoudreza Modarres Aval , Babak Haghshenas
Short-chain fatty acids (SCFAs) are crucial microbial metabolites that mediate host–microbiota interactions, regulate immune responses, and maintain gut homeostasis. While most studies focus on SCFA production by obligate anaerobes, recent evidence highlights Lactobacillus spp. as potential SCFA-producing microorganisms that remain understudied in terms of SCFA-based health and food-related effects. This research review compiles what is currently known about species- and strain-specific biosynthetic capabilities of Lactobacillus for SCFA production, particularly acetate, and lactate, which, while produced by Lactobacillus, is not classified as a SCFA but plays similar metabolic roles (e.g., activation of G-protein-coupled receptors, inhibition of histone deacetylases, and immune cell metabolism).
We additionally assess new synthetic biology and metabolic engineering approaches to improving SCFA yield in Lactobacillus, including gene circuit design, CRISPR editing, and co-culture optimization. Importantly, we discuss translational opportunities not only in inflammatory, metabolic, and neuroimmune diseases but also in the development of functional foods, synbiotics, and nutraceutical applications. Finally, we highlight key challenges—strain variability, delivery strategies, and regulatory oversight—that preclude both clinical and food system translation.
By integrating perspectives from microbiology, immunology, food science, and bioengineering, this review provides a broad framework for the rational design of SCFA-producing Lactobacillus strains as next-generation probiotics and food-grade biotherapeutics.
短链脂肪酸(SCFAs)是重要的微生物代谢物,介导宿主-微生物群相互作用,调节免疫反应,维持肠道稳态。虽然大多数研究都集中在专性厌氧菌生产短链脂肪酸上,但最近的证据表明,乳酸杆菌是潜在的短链脂肪酸生产微生物,但就短链脂肪酸对健康和食品相关的影响而言,其研究仍未充分。本研究综述汇编了目前已知的乳酸杆菌生产SCFA的物种和菌株特异性生物合成能力,特别是醋酸盐和乳酸盐,乳酸虽然由乳酸杆菌产生,但不属于SCFA,但具有类似的代谢作用(例如,激活g蛋白偶联受体,抑制组蛋白去乙酰化酶和免疫细胞代谢)。我们还评估了新的合成生物学和代谢工程方法来提高乳酸杆菌的SCFA产量,包括基因电路设计、CRISPR编辑和共培养优化。重要的是,我们不仅讨论了炎症、代谢和神经免疫疾病的转化机会,还讨论了功能食品、合成生物和营养保健应用的发展机会。最后,我们强调了关键的挑战——菌株变异、递送策略和监管监督——这些都阻碍了临床和食品系统的转化。本文综合了微生物学、免疫学、食品科学和生物工程等方面的观点,为合理设计生产scfa的乳杆菌菌株作为下一代益生菌和食品级生物治疗药物提供了广阔的框架。
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引用次数: 0
Completed genomes from Variovorax provide insight into genome diversification through horizontal gene transfer 来自Variovorax的完整基因组通过水平基因转移提供了基因组多样化的见解
IF 5.8 Q1 MICROBIOLOGY Pub Date : 2025-01-01 DOI: 10.1016/j.crmicr.2025.100497
Christopher J. Ne Ville , Paul M. Orwin
Approximately 10% of all bacterial genomes sequenced thus far contain a secondary replicon. This property of bacterial populations vastly increases genomic diversity within phylogenetically narrow groups. Members of the genus Variovorax have extensive heterogeneity in genome architecture, including sequenced isolates containing plasmids, megaplasmids, and chromids. Many of the Variovorax genomes in the NCBI database were generated using short-read data exclusively and were assembled to the permanent draft stage. We acquired a set of these isolates and used the Oxford Nanopore MinION to generate additional data to allow for hybrid assembly of these genomes. Here we present the finished assemblies of 15 Variovorax isolates from diverse ecosystems that were previously only available as permanent drafts. When added to the previously published Variovorax assemblies for EPS, CSUSB, and VAI-C and those published by other groups, we found significant diversity in genome architecture. We found that there are plasmids, megaplasmids, and chromids that are distinguishable using Guanine-Cytosine (G+C content) content as a signal. We identified a plasmid integration event in NFACC27 and suggest potential evolutionary relationships in the secondary replicons based on ParB homology as well as ANI. The evidence suggests that Variovorax, like its sister taxon Burkholderia, is highly capable of acquiring and maintaining stable secondary replicons. The plasticity of these architectures and the mechanisms for maintenance remain a topic for future research.
到目前为止,大约10%的细菌基因组测序包含一个二级复制子。细菌种群的这种特性极大地增加了系统发育上狭窄群体内的基因组多样性。Variovorax属的成员在基因组结构上具有广泛的异质性,包括含有质粒、巨质粒和染色粒的测序分离物。NCBI数据库中的许多Variovorax基因组都是专门使用短读数据生成的,并被组装到永久草稿阶段。我们获得了一组这些分离物,并使用Oxford Nanopore MinION来生成额外的数据,以便对这些基因组进行杂交组装。在这里,我们展示了来自不同生态系统的15个Variovorax分离物的成品组装,这些分离物以前只能作为永久草稿使用。当将先前发表的针对EPS、CSUSB和VAI-C的Variovorax序列和其他团队发表的序列加在一起时,我们发现了基因组结构的显著多样性。我们发现有质粒、巨质粒和染色粒可以用鸟嘌呤-胞嘧啶(G+C含量)含量作为信号来区分。我们在NFACC27中发现了一个质粒整合事件,并基于ParB同源性和ANI提出了二级复制子之间潜在的进化关系。有证据表明,Variovorax和它的姐妹分类群伯克霍尔德氏菌一样,具有很强的获取和维持稳定的次级复制子的能力。这些结构的可塑性和维持机制仍然是未来研究的主题。
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引用次数: 0
Enhancing diagnostic efficiency of pyrazinamide resistance in Mycobacterium tuberculosis via modified MGIT assay and genotypic correlation 改良MGIT法及基因型相关性提高结核分枝杆菌吡嗪酰胺耐药的诊断效率
IF 5.8 Q1 MICROBIOLOGY Pub Date : 2025-01-01 DOI: 10.1016/j.crmicr.2025.100462
Ananthi Rajendran , Ahmed Kabir Refaya , Balaji Subramanyam , Ramesh Karunaianantham , Dhandapani RaviKumar , Hemalatha Haribabu , Radha Gopalaswamy , Radhika Golla , Vadivel Senthildevi , Narayanan Sivaramakrishnan Gomathi , Sivakumar Shanmugam , Kannan Palaniyandi
Pyrazinamide (PZA) plays a crucial role in the treatment of both active and latent tuberculosis, particularly in regimens designed to treat drug-resistant TB. However, diagnosing resistance to PZA poses challenges for managing TB, highlighting the need for accurate detection methods. This study aims to address the challenges in detecting PZA resistance by modifying the standard MGIT960 PZA drug susceptibility testing method by optimizing the inoculum dilution. Briefly, three MGIT DST versions were evaluated: the standard method, the reduced inoculum (RI) method employing a 1:20 inoculum dilution and the sparse dilution (SD) method using a 1:50 dilution of the inoculum for growth control tube, while the undiluted MGIT positive culture was used for the PZA test tube. The SD MGIT DST approach minimized the number of false-resistant PZA results to (31/401) 7.7 % against 27 % by standard MGIT DST and 11.7 % by RI MGIT DST approach, thereby reducing the false-positivity rate by 19.3 %. Targeted sequencing of pncA gene identified mutations in only 14/401 isolates (3.5 %). Whole genome sequencing (WGS) of the 31 phenotypically resistant isolates identified resistance -associated mutations in pncA gene (45 %), panD (9.6 %), mas (12.9 %), glpK (3.2 %), and lprG (3.2 %), and others efflux associated genes like Rv1258c (3.2 %), Rv0191c (3.2 %), and Rv3008 (6.45 %), except for 4 isolates, for which no mutations were detected in the target genes. These genes are involved in various resistance mechanisms including cell wall synthesis, metabolic pathways, and drug tolerance, which are essential for PZA efficacy. Notably, new mutations in glpK and mas were detected in isolates with wild-type pncA and were absent in the sensitive isolates. Our study substantiates the improvement of phenotypic testing methods and enhances the detection of PZA resistance even in resource-limited settings and direct research towards improving the diagnostic accuracy in TB drug resistance management.
吡嗪酰胺(PZA)在治疗活动性和潜伏性结核病方面发挥着至关重要的作用,特别是在设计用于治疗耐药结核病的方案中。然而,诊断对PZA的耐药性给结核病管理带来了挑战,突出表明需要准确的检测方法。本研究通过优化接种稀释剂,对MGIT960 PZA药敏试验方法进行修改,以解决PZA耐药检测中的难题。简而言之,我们评估了三种MGIT DST版本:标准法,接种量稀释1:20的减少接种量(RI)法和接种量稀释1:50的稀疏稀释(SD)法用于生长对照管,而未稀释的MGIT阳性培养用于PZA试管。SD MGIT DST方法将PZA假抗性结果的数量降至(31/401)7.7%,而标准MGIT DST方法为27%,RI MGIT DST方法为11.7%,从而将假阳性率降低了19.3%。pncA基因的靶向测序仅在14/401株(3.5%)中发现突变。31株表型耐药菌株的全基因组测序(WGS)鉴定出pncA基因(45%)、panD基因(9.6%)、mas基因(12.9%)、glpK基因(3.2%)和lprG基因(3.2%)以及Rv1258c(3.2%)、Rv0191c(3.2%)和rv38(6.45%)等外排相关基因的耐药相关突变,4株菌株靶基因未检测到突变。这些基因参与多种耐药机制,包括细胞壁合成、代谢途径和药物耐受性,这些对PZA的疗效至关重要。值得注意的是,在野生型pncA分离株中检测到glpK和mas的新突变,而在敏感株中则没有。我们的研究证实了表型检测方法的改进,即使在资源有限的情况下也能提高PZA耐药的检测,并直接研究提高结核病耐药管理的诊断准确性。
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引用次数: 0
Quantitative multiplexed analysis of gene and protein expression patterns in Yarrowia lipolytica 脂化耶氏菌基因和蛋白表达模式的定量多重分析
IF 4.8 Q1 MICROBIOLOGY Pub Date : 2025-01-01 DOI: 10.1016/j.crmicr.2025.100369
Erin Bredeweg, Galya Orr, Dehong Hu
In this report, we present coordinated observations of protein and mRNA transcript counts at the single-cell level in the oleaginous yeast model Yarrowia lipolytica. The transcription factor Xbp1p regulates entry into a quiescent state, representing a shift of resources to sequestration of nutrients rather than cell division. We observed the responses of wild-type and Δxbp1 cells to protein (by fluorescence) and transcript quantification and localization at both single-cell and population-averaged levels. Data were collected via single-molecule fluorescence in situ hybridization (smFISH) and qPCR under nitrogen depletion, a condition that drives lipid accumulation. These techniques reveal a complex and heterogeneous population of Xbp1p dynamics and downstream regulation. Our findings highlight the need for single-cell resolution analyses to describe cellular dynamics and regulatory processes.
在本报告中,我们在产油酵母模型多脂耶氏菌的单细胞水平上对蛋白质和mRNA转录计数进行了协调观察。转录因子Xbp1p调节进入静止状态,代表资源转移到隔离营养物质而不是细胞分裂。我们观察了野生型和Δxbp1细胞对蛋白质的反应(通过荧光),并在单细胞和群体平均水平上对转录物进行了定量和定位。数据通过单分子荧光原位杂交(smFISH)和qPCR在氮耗尽(驱动脂质积累的条件)下收集。这些技术揭示了Xbp1p动态和下游调控的复杂和异质群体。我们的研究结果强调需要单细胞分辨率分析来描述细胞动力学和调节过程。
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Current Research in Microbial Sciences
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