Endospore pili: Flexible, stiff, and sticky nanofibers

Jonsmoen, Unni Lise; Malyshev, Dmitry; Berg, R.; Dahlberg, Tobias; Aspholm, Marina; Andersson, Magnus

doi:10.1016/j.bpj.2023.05.024

Cited by 5 publications

(8 citation statements)

References 56 publications

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“…In contrast, the "ladder-like" L-ENAs, are thinner and shorter than the S-ENAs and are linked to the exosporium layer via the spores (Wijman et al, 2007), suggesting that the ENAs may constitute a substantial part of the biofilm matrix. Further research into the S-ENA revealed that they are rigid yet flexible fibers, maintaining integrity under extension, which supports their proposed role in facilitating spore-to-spore binding and potentially enhancing autoaggregation (Jonsmoen et al, 2023). The limited understanding of autoaggregation in B. cereus spp.…”

Section: Introductionmentioning

confidence: 73%

“…The present work builds upon the research conducted by Jonsmoen et al (2023), where single-cell analyses demonstrated the role of S-ENAs in maintaining spores in a gel-like state by mediating distant spore-to-spore interactions (Jonsmoen et al, 2023). Here, by utilizing both bulk and single-cell approaches, we undertook an investigation of the role of L-and S-ENAs, as well as the tip fibrillum of the L-ENAs in the aggregative behavior of B. paranthracis spores, and how different factors in the surrounding liquid influences contacts between spores.…”

Section: Discussionmentioning

confidence: 91%

“…The S++L+ spore pellet was gradually compressed over the days the experiment continued, indicating that the steric hindrance was reduced over time. In Jonsmoen et al, 2023, we demonstrated that spores expressing S-ENA exhibit a gel-like state when suspended in water i.e., a viscosity that indicates relaxed interconnection between spores (Jonsmoen et al, 2023). The gel-like behavior was not observed for ENA-depleted bald spores, which moved independently of each other when suspended in water.…”

Section: Discussionmentioning

confidence: 99%

“…For preparation of spores, the bacteria were streaked onto LB agar plates and incubated for no less than two weeks at 37°C. When approximately 98% of the bacteria had sporulated, as determined by phase contrast microscopy (Olympus BX51, Olympus Corporation, Japan), the spores were harvested as described before in Jonsmoen et al (2023) (Jonsmoen et al, 2023). Pure spore suspensions were stored at 4 °C until the start of the experiment.…”

Section: Methodsmentioning

confidence: 99%

“…The food outbreak isolate B. paranthracis NVH 0075/95 (Lund & Granum, 1996) and the isogenic mutants used in this paper are listed in Table 1 and their phenotype illustrated in Fig 1 . For preparation of spores, the bacteria were streaked onto LB agar plates and incubated for no less than two weeks at 37°C. When approximately 98% of the bacteria had sporulated, as determined by phase contrast microscopy (Olympus BX51, Olympus Corporation, Japan), the spores were harvested as described before in Jonsmoen et al (2023) (Jonsmoen et al, 2023).…”

Section: Strains and Spore Preparationsmentioning

confidence: 99%

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The role of endospore appendages in spore-spore contacts in pathogenic bacilli

Jonsmoen,

Malyshev,

Sleutel

et al. 2024

Preprint

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Species within the spore-forming Bacillus cereus sensu lato group are recognized for their role in food spoilage and food poisoning. B. cereus spores are decorated with numerous pilus-like appendages, called S-ENAs and L-ENAs. These appendages are believed to play crucial roles in self-aggregation, adhesion, and biofilm formation. By using both bulk and single-cell approaches, we investigate the role of S- and L-ENAs as well as the impact of different environmental factors in spore-to-spore contacts and in the interaction between spores and vegetative cells. Our findings reveal that ENAs, and particularly their tip fibrilla, play an essential role in spore self-aggregation but not in the adhesion of spores to vegetative cells. The absence of L-BclA, which builds the L-ENA tip fibrillum, reduced both S- and L-ENA mediated spore aggregation, emphasizing the interconnected roles of S- and L-ENAs. Increased salt concentrations in the liquid environment significantly reduced spore aggregation, implying a charge dependency of spore-spore interactions. By elucidating these complex interactions, our study provides valuable insights into spore dynamics. This knowledge can guide future studies on spore behavior in environmental settings and aids in developing strategies to manage bacterial aggregation for beneficial purposes, like controlling biofilms in food production equipment.

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Section: Introductionmentioning

confidence: 73%

Section: Discussionmentioning

confidence: 91%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Strains and Spore Preparationsmentioning

confidence: 99%

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The role of endospore appendages in spore-spore contacts in pathogenic bacilli

Jonsmoen,

Malyshev,

Sleutel

et al. 2024

Preprint

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Evaluating Bacterial Spore Preparation Methods for Scanning Electron Microscopy

Malyshev,

Lee,

Andersson

2024

Microscopy and Microanalysis

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Scanning electron microscopy (SEM) can reveal the ultrastructure of bacterial spores, including morphology, surface features, texture, spore damage, germination, and appendages. Understanding these features can provide a basis for adherence, how physical and environmental stressors affect spore viability, integrity, and functionality, as well as the distribution and function of surface appendages. However, the spore sample preparation method can significantly impact the SEM images’ appearance, resolution, and overall quality. In this study, we compare different spore preparation methods to identify optimal approaches for preparation time, spore appearance and resolved features, including the exosporium and spore pili, for SEM imaging. We use Bacillus paranthracis as model species and evaluate the efficacy of preparation protocols using different fixation and drying methods, as well as imaging under room- and cryogenic temperatures. We compare and assess method complexity to the visibility of the spore exosporium and spore appendages across different methods. Additionally, we use Haralick texture features to quantify the differences in spore surface appearance and determine the most suitable method for preserving spore structures and surface features during SEM evaluation. The findings from this study will help establish protocols for preparing bacterial spores for SEM and facilitating accurate and reliable analysis of spores’ characteristics.

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Physico-chemical characterization of single bacteria and spores using optical tweezers

Nilsson

Jonsmoen

Malyshev

et al. 2022

Preprint

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Spore-forming pathogenic bacteria are adapted for adhering to surfaces, and their endospores can tolerate strong chemicals making decontamination difficult. Understanding the physico-chemical properties of bacteria and spores is therefore essential in developing antiadhesive surfaces and disinfection techniques. However, measuring physico-chemical properties in bulk does not show the heterogeneity between cells. Characterizing bacteria on a single-cell level can provide mechanistic clues usually hidden in bulk measurements. This paper shows how optical tweezers can be applied to characterize single bacteria and spores, and how physico-chemical properties related to adhesion, fluid dynamics, biochemistry, and metabolic activity can be assessed.

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Endospore pili: Flexible, stiff, and sticky nanofibers

Cited by 5 publications

References 56 publications

The role of endospore appendages in spore-spore contacts in pathogenic bacilli

The role of endospore appendages in spore-spore contacts in pathogenic bacilli

Evaluating Bacterial Spore Preparation Methods for Scanning Electron Microscopy

Physico-chemical characterization of single bacteria and spores using optical tweezers

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