IF 5.9 Q1 MICROBIOLOGY

Biofilm

Pub Date : 2025-04-15 DOI: 10.1016/j.bioflm.2025.100280

Aaron Crowther , Gareth LuTheryn , Ramón Garcia-Maset , Maryam Parhizkar , J. Mark Sutton , Charlotte Hind , Dario Carugo

Opportunistic pathogen Pseudomonas aeruginosa plays a crucial role in chronic wound biofilms, increasing infection's morbidity and mortality. In recent years, the signalling molecule nitric oxide (NO) and chelating agent tetrasodium EDTA (T-EDTA) have been applied therapeutically owing to their multifactorial effects including bacterial killing, biofilm dispersal, and wound healing. However, previous studies assessing NO's antibiofilm efficacy have not considered the variable pH and temperature of the wound environment. Here, pH-dependent NO donors N-diazeniumdiolates (NONOates), PAPA NONOate (PA-NO) and Spermine NONOate (SP–NO), and T-EDTA were applied in wound-relevant pH environments (pH 5.5–8.5) and temperatures (32 °C and 37 °C) to P. aeruginosa PAO1 biofilms grown for either 24 or 48 h. At 32 °C and pH 7.5, 250 μM PA-NO reduced 24-h biofilm biomass by 35 %. At 37 °C, 250 μM PA-NO and 4 % w/v T-EDTA caused 21 % and 57 % biomass reduction in 24-h biofilms, respectively. In 48-h biofilms, NONOates did not induce significant biomass reduction, while T-EDTA maintained its efficacy with a 64 % reduction. A subsequent experiment investigated the impact of NONOates and T-EDTA as pre-treatments before exposure to ciprofloxacin. Unexpectedly, NONOate pre-treatment decreased ciprofloxacin's effectiveness, resulting in approximately 1-log increase in viable planktonic and biofilm-residing cells compared to ciprofloxacin alone. It was hypothesized that this protective effect might stem from NO-induced decreased cellular respiration, which inhibits reactive oxygen species (ROS)-mediated bactericidal mechanisms. These findings highlight both the potential and complexities of developing effective antimicrobial strategies for chronic wound infections, emphasizing the need for further research to optimize treatment approaches.

机会致病菌铜绿假单胞菌在慢性伤口生物膜中起着至关重要的作用，增加了感染的发病率和死亡率。近年来，信号分子一氧化氮（NO）和螯合剂四钠EDTA （T-EDTA）由于其多因素作用，包括细菌杀灭、生物膜分散和伤口愈合，已被应用于治疗。然而，以往评估NO抗生素膜功效的研究没有考虑到伤口环境的pH和温度的变化。在这里，pH依赖性NO供体n -重氮双酸酯（NONOates）， PAPA NONOate （PA-NO）和精胺NONOate （SP-NO）以及T-EDTA在伤口相关的pH环境（pH 5.5-8.5）和温度（32°C和37°C）下对铜绿假单胞菌PAO1生物膜生长24或48小时。在32°C和pH 7.5下，250 μM PA-NO使24小时的生物膜生物量减少35%。在37°C时，250 μM PA-NO和4% w/v T-EDTA分别使24 h生物膜的生物量减少21%和57%。在48 h的生物膜中，NONOates没有引起显著的生物量减少，而T-EDTA保持其效果，减少了64%。随后的实验研究了NONOates和T-EDTA作为暴露于环丙沙星前的预处理的影响。出乎意料的是，NONOate预处理降低了环丙沙星的有效性，与单独环丙沙星相比，导致浮游生物和生物膜存活细胞增加了大约1倍。据推测，这种保护作用可能源于no诱导的细胞呼吸减少，从而抑制活性氧（ROS）介导的杀菌机制。这些发现强调了开发有效的抗微生物策略治疗慢性伤口感染的潜力和复杂性，强调需要进一步研究以优化治疗方法。

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引用次数: 0

Modeling reciprocal adaptation of Staphylococcus aureus and Pseudomonas aeruginosa co-isolates in artificial sputum medium 模拟金黄色葡萄球菌和铜绿假单胞菌共分离物在人工痰液中的相互适应

IF 5.9 Q1 MICROBIOLOGY

Biofilm

Pub Date : 2025-04-11 DOI: 10.1016/j.bioflm.2025.100279

Zhifen Wang , Emily Giedraitis , Christiane Knoop , Daniel J. Breiner , Vanessa V. Phelan , Françoise Van Bambeke

Co-infections by Staphylococcus aureus and Pseudomonas aeruginosa are frequent in the airways of patients with cystic fibrosis. These co-infections show higher antibiotic tolerance in vitro compared to mono-infections. In vitro models have been developed to study the interspecies interactions between P. aeruginosa and S. aureus. However, these model systems fail to incorporate clinical isolates with diverse phenotypes, do not reflect the nutritional environment of the CF airway mucus, and/or do not model the biofilm mode of growth observed in the CF airways. Here, we established a dual-species biofilm model grown in artificial sputum medium, where S. aureus was inoculated before P. aeruginosa to facilitate the maintenance of both species over time. It was successfully applied to ten pairs of clinical isolates exhibiting different phenotypes. Co-isolates from individual patients led to robust, stable co-cultures, supporting the theory of cross-adaptation in vivo. Investigation into the cross-adaptation of the VBB496 co-isolate pair revealed that both the P. aeruginosa and S. aureus isolates had reduced antagonism, in part due to reduced production of P. aeruginosa secondary metabolites as well as higher tolerance to those metabolites by S. aureus. Together, these results indicate that the two-species biofilm model system provides a useful tool for exploring interspecies interactions of P. aeruginosa and S. aureus in the context of CF airway infections.

金黄色葡萄球菌和铜绿假单胞菌的合并感染在囊性纤维化患者的气道中是常见的。与单一感染相比，这些合并感染在体外表现出更高的抗生素耐受性。为了研究铜绿假单胞菌和金黄色葡萄球菌之间的种间相互作用，已经建立了体外模型。然而，这些模型系统未能纳入具有不同表型的临床分离株，不能反映CF气道粘液的营养环境，和/或不能模拟CF气道中观察到的生物膜生长模式。在这里，我们建立了一个在人工痰培养基中培养的双物种生物膜模型，在铜绿假单胞菌之前接种金黄色葡萄球菌，以促进两种物种的长期维持。该方法成功应用于表现不同表型的10对临床分离株。从个体患者中分离出的共培养产生了强大、稳定的共培养，支持了体内交叉适应理论。对VBB496共分离物对的交叉适应研究表明，铜绿假单胞菌和金黄色葡萄球菌分离物的拮抗作用都降低了，部分原因是铜绿假单胞菌次生代谢物的产生减少，以及金黄色葡萄球菌对这些代谢物的耐受性提高。总之，这些结果表明，两种生物膜模型系统为探索铜绿假单胞菌和金黄色葡萄球菌在CF气道感染中的种间相互作用提供了有用的工具。

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引用次数: 0

The burden of antimicrobial resistance in biofilm forming Staphylococcus spp. from Vernal Keratoconjunctivitis patients eyes 春性角膜结膜炎患者眼部生物膜形成葡萄球菌的耐药性负担

IF 5.9 Q1 MICROBIOLOGY

Biofilm

Pub Date : 2025-04-05 DOI: 10.1016/j.bioflm.2025.100278

Nelaveni Rupa , Pragnya Rao Donthineni , Sayan Basu , Kotakonda Arunasri

Vernal keratoconjunctivitis (VKC) is a chronic allergic ocular surface disease with seasonal recurrences and severe forms showing vision threatening complications. The purpose of the study is to understand the prevalence and diversity of biofilm-forming bacteria and antimicrobial resistance in VKC compared to healthy individuals (HC). For this, conjunctival swab samples were collected from VKC (n = 26) and HC (n = 23), of which culture positive samples were 77 % and 78.26 % respectively. The 16S rRNA gene sequencing revealed a significant increase in bacterial diversity in VKC compared to HC (p < 0.05), identifying 16 and 9 bacterial species, respectively. Staphylococcus epidermidis emerged as the predominant bacterium in both groups, with relative abundances of 52.8 % in HC and 30.2 % in VKC (p < 0.001). Biofilm formation was observed in 64.15 % of bacterial species in VKC and 31 % in HC (p < 0.001). Scanning electron microscopy analysis confirmed temporal biofilm formation by Staphylococcus spp. in both groups. Minimum inhibitory concentration testing showed that biofilm forming Staphylococcus spp. from VKC exhibited multidrug resistance (>2 antibiotics) more frequently than those from HC. Additionally, Staphylococcus spp. in VKC demonstrated higher resistance to fluoroquinolones compared to HC. These findings indicate a significantly greater prevalence of biofilm-forming and antimicrobial resistant Staphylococcus bacteria in VKC Patients compared with HC.

春性角膜结膜炎（VKC）是一种慢性过敏性眼表疾病，季节性复发，严重形式表现为威胁视力的并发症。本研究的目的是了解与健康个体（HC）相比，VKC中生物膜形成细菌的患病率和多样性以及抗菌素耐药性。为此，收集VKC （n = 26）和HC （n = 23）结膜拭子样本，其中培养阳性样本分别为77%和78.26%。16S rRNA基因测序显示，与HC相比，VKC中细菌多样性显著增加(p <；0.05)，分别鉴定出16种和9种细菌。表皮葡萄球菌在两组中均为优势菌，在HC和VKC中相对丰度分别为52.8%和30.2% (p <；0.001)。64.15%的VKC细菌和31%的HC细菌形成生物膜(p <；0.001)。扫描电镜分析证实两组均有葡萄球菌形成的时间生物膜。最低抑菌浓度试验表明，VKC产生物膜葡萄球菌比HC产多药耐药（2种抗生素）的频率更高。此外，与HC相比，VKC中的葡萄球菌对氟喹诺酮类药物具有更高的耐药性。这些发现表明，与HC相比，VKC患者中生物膜形成和抗微生物葡萄球菌的患病率明显更高。

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引用次数: 0

Diverse reactions of aquaculture biofilter biofilms following acute high-dose peracetic acid 急性高剂量过氧乙酸后水产养殖生物滤池生物膜的多种反应

IF 5.9 Q1 MICROBIOLOGY

Biofilm

Pub Date : 2025-04-02 DOI: 10.1016/j.bioflm.2025.100277

Wanhe Qi , Sanni L. Aalto , Peter Vilhelm Skov , Kim João de Jesus Gregersen , Lars-Flemming Pedersen

Peracetic acid (PAA) is an effective disinfectant in aquaculture systems to reduce pathogen loads and improve water quality. However, its effectiveness in disinfecting biofilm in recirculating aquaculture systems (RAS) and resetting biofilters between productions remains unknown. This study evaluated the effects of acute PAA exposure on biofilter biofilms from freshwater RAS. Identical types of bioelements were collected from a pilot-scale RAS (without prior PAA treatment) and a commercial RAS (with PAA treatment), and exposed to PAA concentrations of 0, 1, 2, 4, 8, and 16 mg/L for 1 h. Microbial activity and viability of the exposed biofilms were evaluated using respirometry and flow cytometry. Results showed dose-dependent inhibition of biofilm activity and viability in the pilot-scale RAS. Nitrite oxidation was the most sensitive process to PAA, with an IC₅₀ of 1.27 mg/L (the concentration at which PAA inhibited biofilm metabolic activity by 50 %), followed by ammonia oxidation (IC₅₀ = 1.59 mg/L) and endogenous respiration (IC₅₀ = 2.67 mg/L). Microbial activity linked to H₂O₂ decomposition was least affected (IC₅₀ = 4.68 mg/L). Live cell counts decreased from 9.1 × 10⁷ counts/cm² to 2.4 × 10⁷ counts/cm² of bioelement surface, with dead cells proportion increasing from 15 % to 54 %. In contrast, biofilter biofilms from the commercial RAS exhibited significantly lower sensitivity to PAA dosage, with reductions in nitrite oxidation (39 %) and ammonia oxidation (51 %) observed only at 16 mg/L compared to control. These findings suggest that routine PAA exposure, as part of the other operating conditions on the commercial RAS, can enhance the biofilm's sensitivity to PAA. The study provides new insight into the sensitivity of aquaculture biofilm to PAA treatment and its effect on associated microbial processes.

过氧乙酸（PAA）是水产养殖系统中降低病原菌负荷、改善水质的有效消毒剂。然而，它在循环水养殖系统（RAS）中消毒生物膜和在生产之间重置生物过滤器方面的有效性仍然未知。本研究评估急性PAA暴露对淡水RAS生物过滤器生物膜的影响。从中试RAS（未经事先PAA处理）和商用RAS（经PAA处理）中收集相同类型的生物元素，并将其暴露于浓度为0、1、2、4、8和16 mg/L的PAA中1小时。使用呼吸仪和流式细胞术评估暴露的生物膜的微生物活性和活力。结果显示，中试RAS对生物膜活性和活力的抑制呈剂量依赖性。亚硝酸盐氧化对PAA最敏感，其IC50为1.27 mg/L （PAA抑制生物膜代谢活性50%的浓度），其次是氨氧化（IC50 = 1.59 mg/L）和内源呼吸（IC50 = 2.67 mg/L）。与H2O2分解相关的微生物活性受影响最小（IC50 = 4.68 mg/L）。活细胞计数从9.1 × 107个/cm2下降到2.4 × 107个/cm2，死亡细胞比例从15%上升到54%。相比之下，来自商业RAS的生物过滤器生物膜对PAA剂量的敏感性显着降低，与对照相比，仅在16 mg/L时观察到亚硝酸盐氧化（39%）和氨氧化（51%）的减少。这些发现表明，常规的PAA暴露，作为商业RAS的其他操作条件的一部分，可以增强生物膜对PAA的敏感性。该研究为水产养殖生物膜对PAA处理的敏感性及其对相关微生物过程的影响提供了新的见解。

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引用次数: 0

Bacterial cellulose: Enhancing productivity and material properties through repeated harvest 细菌纤维素：通过反复收获提高生产率和材料性能

IF 5.9 Q1 MICROBIOLOGY

Biofilm

Pub Date : 2025-03-26 DOI: 10.1016/j.bioflm.2025.100276

N. Rackov , N. Janßen , A. Akkache , B. Drotleff , B. Beyer , E. Scoppola , N.E. Vrana , R. Hengge , C.M. Bidan , S. Hathroubi

Bacterial cellulose (BC), a promising versatile biopolymer produced by bacteria, has an immense potential in various industries. However, large-scale application is hindered by high production costs and low yields. This study introduces an innovative approach combining a prolonged static culturing with intermittent harvesting. This novel strategy resulted in a significant increase in BC productivity, achieving up to a threefold rise in biomass within the first 35 days. Prolonged growth and continuous harvesting not only enhanced productivity but also led to a mutant strain M2 with higher yields and distinct BC architecture. Mechanical and structural analyses revealed that sequential harvest correlated with increasing crystallinity, altered crystallite sizes, and improved stiffness of the dry material during initial cycles, potentially reflecting bacteria adaptation to resources limitations. Genomic analysis identified key mutations in the M2 strain, including one in the RelA/SpoT enzyme, suggesting a reduced stringent response that promotes growth under nutrient-limiting conditions. Untargeted metabolomic profiling revealed deregulation of several metabolites, including a significant difference in fatty acid metabolites that could potentially influence membrane fluidity and BC secretion. Such metabolic and structural adaptations enhance BC production efficiency and material properties. These findings highlight the potential of intermittent harvesting for sustainable BC production and the role of bacterial adaptation in tuning BC properties. Further research will optimize this strategy and expand its applications in developing tailored biomaterials for diverse industries.

细菌纤维素（BC）是一种很有前途的由细菌产生的多用途生物聚合物，在各个工业领域具有巨大的潜力。然而，高生产成本和低产量阻碍了大规模应用。本研究介绍了一种将长时间静态培养与间歇收获相结合的创新方法。这种新颖的策略导致了BC生产力的显著增加，在前35天内实现了生物量的三倍增长。长时间生长和连续收获不仅提高了产量，而且产生了产量更高、BC结构独特的突变株M2。机械和结构分析显示，在初始循环中，连续收获与结晶度增加、晶体大小改变和干燥材料刚度提高相关，这可能反映了细菌对资源限制的适应。基因组分析确定了M2菌株的关键突变，包括RelA/SpoT酶的一个突变，表明在营养限制条件下促进生长的严格反应降低。非靶向代谢组学分析揭示了几种代谢物的失调，包括脂肪酸代谢物的显著差异，这可能会影响膜流动性和BC分泌。这种代谢和结构适应提高了BC的生产效率和材料性能。这些发现强调了间歇采收对可持续BC生产的潜力以及细菌适应在调整BC特性中的作用。进一步的研究将优化这一策略，并扩大其在开发不同行业定制生物材料方面的应用。

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引用次数: 0

Nanowire arrays with programmable geometries as a highly effective anti-biofilm surface 具有可编程几何形状的纳米线阵列作为一种高效的抗生物膜表面

IF 5.9 Q1 MICROBIOLOGY

Biofilm

Pub Date : 2025-03-24 DOI: 10.1016/j.bioflm.2025.100275

Marina A. George , David McGiffin , Anton Y. Peleg , Roey Elnathan , David M. Kaye , Yue Qu , Nicolas H. Voelcker

Biofilm-related microbial infections are the Achilles’ heel of many implantable medical devices. Surface patterning with nanostructures in the form of vertically aligned silicon (Si) nanowires (VA-SiNWs) holds promise to prevent these often “incurable” infections. In this study, we fabricated arrays of highly ordered SiNWs varying in three geometric parameters, including height, pitch size, and tip diameter (sharpness). Anti-infective efficacies of fabricated SiNW arrays were assessed against representative laboratory reference bacterial strains, Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922, using a modified microwell biofilm assay representing microorganism-implant interactions at a liquid-solid interface. To further understand the role of individual geometric parameters to the SiNW-induced bacterial killing, SiNW arrays with stepwise changes in individual geometric parameters were compared. The force that NWs applied on bacterial cells was mathematically calculated. Our results suggested that NWs with specific geometries were able to kill adherent bacterial cells and prevent further biofilm formation on biomaterial surfaces. Tip diameter and pitch size appeared to be key factors of nanowires predetermining their anti-infectiveness. Mechanistic investigation found that tip diameter and pitch size co-determined the pressure that NWs put on the cell envelope. The most effective anti-infective NWs fabricated in our study (50 nm in tip diameter and 400 nm in pitch size for S. aureus and 50 nm in tip diameter and 800 nm in pitch size for E. coli) put pressures of approximately 2.79 Pa and 8.86 Pa to the cell envelop of S. aureus and E. coli, respectively, and induced cell lyses. In addition, these NWs retained their activities against clinical isolates of S. aureus and E. coli from patients with confirmed device-related infections and showed little toxicity against human fibroblast cells and red blood cells.

生物膜相关的微生物感染是许多植入式医疗器械的致命弱点。以垂直排列的硅（Si）纳米线（VA-SiNWs）形式的纳米结构的表面图案有望预防这些通常“无法治愈”的感染。在这项研究中，我们制造了高度有序的sinw阵列，其几何参数包括高度、节距大小和尖端直径（锐度）。采用改进的微孔生物膜法，对制备的SiNW阵列对代表性实验室参考菌株金黄色葡萄球菌（Staphylococcus aureus） ATCC 25923和大肠杆菌（Escherichia coli） ATCC 25922的抗感染效果进行了评估，该方法代表了微生物与植入物在液固界面的相互作用。为了进一步了解单个几何参数对SiNW诱导细菌杀灭的作用，我们比较了单个几何参数逐步变化的SiNW阵列。用数学方法计算了NWs对细菌细胞施加的力。我们的研究结果表明，具有特定几何形状的NWs能够杀死附着的细菌细胞，并阻止生物材料表面进一步形成生物膜。针尖直径和节距大小是决定纳米线抗感染能力的关键因素。机制研究发现，尖端直径和节距大小共同决定了NWs对细胞包膜施加的压力。本研究制备的最有效的抗感染NWs（金黄色葡萄球菌的尖端直径为50 nm，间距为400 nm，大肠杆菌的尖端直径为50 nm，间距为800 nm）分别对金黄色葡萄球菌和大肠杆菌的细胞膜施加约2.79 Pa和8.86 Pa的压力，并诱导细胞裂解。此外，这些NWs保留了对临床分离的金黄色葡萄球菌和大肠杆菌的活性，这些临床分离的金黄色葡萄球菌和大肠杆菌来自确诊的器械相关感染的患者，并且对人成纤维细胞和红细胞的毒性很小。

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引用次数: 0

Decoding the impact of interspecies interactions on biofilm matrix components 解码物种间相互作用对生物膜基质成分的影响

IF 5.9 Q1 MICROBIOLOGY

Biofilm

Pub Date : 2025-03-14 DOI: 10.1016/j.bioflm.2025.100271

Cristina I. Amador , Henriette L. Røder , Jakob Herschend , Thomas R. Neu , Mette Burmølle

Multispecies biofilms are complex communities where extracellular polymeric substances (EPS) shape structure, adaptability, and functionality. However, characterizing the components of EPS, particularly glycans and proteins, remains a challenge due to the diverse bacterial species present and their interactions within the matrix. This study examined how interactions between different species affect EPS component composition and spatial organization. We analyzed a consortium of four bacterial soil isolates that have previously demonstrated various intrinsic properties in biofilm communities: Microbacterium oxydans, Paenibacillus amylolyticus, Stenotrophomonas rhizophila, and Xanthomonas retroflexus. We used fluorescence lectin binding analysis to identify specific glycan components and meta-proteomics to characterize matrix proteins in mono- and multispecies biofilms. Our results revealed diverse glycan structures and composition, including fucose and different amino sugar-containing polymers, with substantial differences between monospecies and multispecies biofilms. In isolation, M. oxydans produced galactose/N-Acetylgalactosamine network-like structures and influenced the matrix composition in multispecies biofilms. Proteomic analysis revealed presence of flagellin proteins in X. retroflexus and P. amylolyticus, particularly in multispecies biofilms. Additionally, surface-layer proteins and a unique peroxidase were identified in P. amylolyticus multispecies biofilms, indicating enhanced oxidative stress resistance and structural stability under these conditions. This study highlights the crucial role of interspecies interactions in shaping the biofilm matrix, as well as the production of glycans and proteins, thereby enhancing our understanding of biofilm complexity.

多物种生物膜是细胞外聚合物质（EPS）形成结构、适应性和功能的复杂群落。然而，表征EPS的成分，特别是聚糖和蛋白质，仍然是一个挑战，因为存在不同的细菌种类和它们在基质内的相互作用。本研究考察了不同物种间相互作用对EPS成分组成和空间组织的影响。我们分析了四种细菌土壤分离物的联盟，这些细菌以前在生物膜群落中表现出各种内在特性：氧化微杆菌、解淀粉芽孢杆菌、嗜根寡养单胞菌和逆转录黄单胞菌。我们使用荧光凝集素结合分析来鉴定特定的聚糖组分，并使用元蛋白质组学来表征单物种和多物种生物膜中的基质蛋白。我们的研究结果揭示了不同的聚糖结构和组成，包括焦点和不同的氨基糖含聚合物，在单物种和多物种生物膜之间存在实质性差异。在分离条件下，M. oxydans产生半乳糖/ n -乙酰半乳糖胺网状结构，并影响多物种生物膜的基质组成。蛋白质组学分析显示，在逆行X.和解淀粉X.中存在鞭毛蛋白，特别是在多物种生物膜中。此外，在P. amylolyticus多物种生物膜中发现了表面层蛋白和独特的过氧化物酶，表明在这些条件下，P. amylolyticus生物膜的抗氧化性和结构稳定性增强。这项研究强调了物种间相互作用在形成生物膜基质中的关键作用，以及聚糖和蛋白质的产生，从而增强了我们对生物膜复杂性的理解。

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引用次数: 0

The biofilm matrix protects Bacillu subtilis against hydrogen peroxide 生物膜基质保护枯草芽孢杆菌免受过氧化氢的侵害

IF 5.9 Q1 MICROBIOLOGY

Biofilm

Pub Date : 2025-03-13 DOI: 10.1016/j.bioflm.2025.100274

Erika Muratov , Julian Keilholz , Ákos T. Kovács , Ralf Moeller

Biofilms formed by Bacillus subtilis confer protection against environmental stressors through extracellular polysaccharides (EPS) and sporulation. This study investigates the roles of these biofilm components in resistance to hydrogen peroxide, a common reactive oxygen species source and disinfectant. Using wild-type and mutant strains deficient in EPS or sporulation, biofilm colonies were cultivated at various maturation stages and exposed to hydrogen peroxide. EPS-deficient biofilms exhibited reduced resilience, particularly in early stages, highlighting the structural and protective importance of the matrix. Mature biofilms demonstrated additional protective mechanisms, potentially involving TasA protein fibers and/or the biofilm surface layer (BslA). In contrast, sporulation showed limited contribution to hydrogen peroxide resistance, as survival was primarily matrix-dependent. These findings underscore the necessity of targeting EPS and other matrix components in anti-biofilm strategies, suggesting that hydrogen peroxide-based disinfection could be enhanced by combining it with complementary sporicidal treatments. This study advances our understanding of biofilm resilience, contributing to the development of more effective sterilization protocols.

由枯草芽孢杆菌形成的生物膜通过胞外多糖（EPS）和孢子形成，对环境胁迫具有保护作用。本研究探讨了这些生物膜成分在抵抗过氧化氢（一种常见的活性氧源和消毒剂）中的作用。利用缺乏EPS或产孢的野生型和突变型菌株，在不同的成熟阶段培养生物膜菌落并暴露于过氧化氢中。缺乏eps的生物膜表现出较低的弹性，特别是在早期阶段，突出了基质的结构和保护重要性。成熟的生物膜表现出额外的保护机制，可能涉及TasA蛋白纤维和/或生物膜表面层（BslA）。相比之下，孢子形成对过氧化氢抗性的贡献有限，因为存活主要依赖于基质。这些发现强调了在抗生物膜策略中靶向EPS和其他基质成分的必要性，这表明基于过氧化氢的消毒可以通过将其与补充的杀孢剂治疗相结合来增强。这项研究促进了我们对生物膜弹性的理解，有助于开发更有效的灭菌方案。

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引用次数: 0

A biofilm-targeting lipo-peptoid to treat Pseudomonas aeruginosa and Staphylococcus aureus co-infections 一种生物膜靶向脂肽类药物治疗铜绿假单胞菌和金黄色葡萄球菌合并感染

IF 5.9 Q1 MICROBIOLOGY

Biofilm

Pub Date : 2025-03-12 DOI: 10.1016/j.bioflm.2025.100272

Samuel J.T. Wardell , Deborah B.Y. Yung , Josefine E. Nielsen , Rajesh Lamichhane , Kristian Sørensen , Natalia Molchanova , Claudine Herlan , Jennifer S. Lin , Stefan Bräse , Lyn M. Wise , Annelise E. Barron , Daniel Pletzer

Antibiotic-resistant bacterial infections are a significant clinical challenge, especially when involving multiple species. Antimicrobial peptides and their synthetic analogues, peptoids, which target bacterial cell membranes as well as intracellular components, offer potential solutions. We evaluated the biological activities of novel peptoids TM11-TM20, which include an additional charged NLys residue, against multidrug-resistant Pseudomonas aeruginosa and Staphylococcus aureus, both in vitro and in vivo. Building on insights from previously reported compounds TM1-TM10, the lipo-peptoid TM18, which forms self-assembled ellipsoidal micelles, demonstrated potent antimicrobial, anti-biofilm, and anti-abscess activity. Transcriptome sequencing (RNA-seq) revealed that TM18 disrupted gene expression pathways linked to antibiotic resistance and tolerance, and biofilm formation in both pathogens. Under dual-species conditions, TM18 induced overlapping but attenuated transcriptional changes, suggesting a priming effect that enhances bacterial tolerance. In a murine skin infection model, TM18 significantly reduced dermonecrosis and bacterial burden in mono-species infections. When combined with the antibiotic meropenem, they synergistically nearly cleared co-infections. Our findings highlight that TM18 has potential as a novel therapeutic for combating antibiotic-resistant pathogens and associated biofilm-driven tolerance.

耐抗生素细菌感染是一个重大的临床挑战，特别是涉及多个物种时。抗菌肽及其合成类似物，类肽，以细菌细胞膜和细胞内成分为目标，提供了潜在的解决方案。我们在体外和体内评估了新型肽类TM11-TM20对多重耐药铜绿假单胞菌和金黄色葡萄球菌的生物活性，TM11-TM20含有一个额外的带电荷的nys残基。基于先前报道的化合物TM1-TM10的见解，脂肽类TM18形成自组装椭球状胶束，显示出有效的抗菌，抗生物膜和抗脓肿活性。转录组测序（RNA-seq）显示，TM18破坏了两种病原体中与抗生素耐药性和耐受性以及生物膜形成相关的基因表达途径。在双种条件下，TM18诱导重叠但减弱转录变化，表明启动效应增强了细菌的耐受性。在小鼠皮肤感染模型中，TM18显著降低了单物种感染的皮肤坏死和细菌负担。当与抗生素美罗培南联合使用时，它们协同作用几乎清除了合并感染。我们的研究结果强调，TM18有潜力作为对抗抗生素耐药病原体和相关生物膜驱动耐受性的新疗法。

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引用次数: 0

Strain-dependent emergence of aminoglycoside resistance in Escherichia coli biofilms 大肠埃希菌生物膜中氨基糖苷类药物耐药性的出现取决于菌株

IF 5.9 Q1 MICROBIOLOGY

Biofilm

Pub Date : 2025-03-12 DOI: 10.1016/j.bioflm.2025.100273

Raphaël Charron , Pierre Lemée , Antoine Huguet , Ornella Minlong , Marine Boulanger , Paméla Houée , Christophe Soumet , Romain Briandet , Arnaud Bridier

In most Earth environments, bacteria predominantly exist within surface-associated communities known as biofilms, where they are embedded in an extracellular matrix. These collective structures play a critical role in bacterial physiology and significantly shape their evolutionary trajectories, contributing to the development of antimicrobial resistance and enhancing bacterial resilience to treatments, with profound implications for public health. This study assessed the impact of the biofilm lifestyle on the emergence of resistance to gentamicin, an aminoglycoside antibiotic, in one laboratory reference strain and seven Escherichia coli isolates from food-processing environments. Throughout a one-month evolution experiment, we observed that certain strains showed a markedly higher emergence of gentamicin-resistant variants in biofilms than in planktonic states, with the emergence of stable variants being closely linked to biofilm maturation. Genomic and phenotypic analyses of gentamicin-resistant (GenR) variants uncovered varied adaptive strategies among the strains. GenR variants from two food-processing isolates (Ec709 and Ec478) displayed point mutations in genes associated with central carbon metabolism (aceE, ygfZ, …) and cell respiration (atpG, cydA, …), while retaining relative growth and colonization capacities. Conversely, GenR variants from the reference strain (Ec1655) adapted preferentially through large genomic deletions, including consistent loss of the peptide transporter gene sbmA, significantly altering cellular fitness. These findings highlight the complexity of adaptive evolution in biofilms and underscore the importance of investigating diverse strains to grasp the full spectrum of adaptation in natural bacterial populations.

在大多数地球环境中，细菌主要存在于被称为生物膜的表面相关群落中，它们被嵌入细胞外基质中。这些集体结构在细菌生理学中发挥关键作用，并显著塑造其进化轨迹，促进抗菌素耐药性的发展，增强细菌对治疗的适应能力，对公共卫生产生深远影响。本研究评估了生物膜生活方式对一种实验室参考菌株和7种食品加工环境中分离的大肠杆菌对庆大霉素（一种氨基糖苷类抗生素）产生耐药性的影响。在为期一个月的进化实验中，我们观察到某些菌株在生物膜中出现的庆大霉素抗性变体明显高于浮游状态，稳定变体的出现与生物膜成熟密切相关。庆大霉素耐药（GenR）变异的基因组和表型分析揭示了菌株之间不同的适应策略。来自两个食品加工分离株（Ec709和Ec478）的GenR变异在与中心碳代谢（aceE, ygfZ，…）和细胞呼吸（atpG, cydA，…）相关的基因上显示点突变，同时保持相对生长和定植能力。相反，参考菌株（Ec1655）的GenR变体优先适应大的基因组缺失，包括肽转运基因sbmA的持续缺失，显著改变了细胞适应性。这些发现突出了生物膜适应进化的复杂性，并强调了研究不同菌株以掌握天然细菌群体适应的全部范围的重要性。

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引用次数: 0

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