Quiescent Body as a Reversible Subcellular Structure Formed in Nongrowing Bacterial Cells

2018 ◽  
Author(s):  
Jiayu Yu ◽  
Yang Liu ◽  
Huijia Yin ◽  
Zengyi Chang
Keyword(s):  
Bacterial Cells ◽  
Subcellular Structure

scholarly journals Quiescent Body as a Reversible Subcellular Structure Formed in Non-growing Bacterial Cells

2017 ◽  
Author(s):  
Jiayu Yu ◽  
Yang Liu ◽  
Huijia Yin ◽  
Zengyi Chang
Keyword(s):  
Cell Growth ◽  
Bacterial Growth ◽  
Lag Time ◽  
Cellular Respiration ◽  
Antibiotic Tolerance ◽  
Bacterial Cells ◽  
Subcellular Structure ◽  
Time Duration ◽  
Highly Correlated

SummaryBacterial cells take a variable lag time and maintain a multi-drug tolerant non-growing state before resuming growth when encountering growth-supportive conditions. Some of them exhibit an extraordinarily long lag, as pathogenic persisters do. It remains unknown on what determines lag time duration. Here, we unveiled a subcellular structure, termed quiescent body, that is formed in bacterial cells entering non-growing state and sequesters selected proteins essential for cell growth. Their formation occurs progressively in each cell and heterogeneously among individual cells. They only dissolve in re-growing cells to release proteins for immediate re-functioning when conditions become fit. Quiescent body, whose degree of formation is highly correlated with duration of lag time or level of antibiotic tolerance, apparently functions as a biological timer for bacterial growth resumption. Further, suppressing their formation, which directly relies on cellular respiration complexes, or promoting their dissolution might be a viable strategy to eradicate persisters.

scholarly journals Regrowth-delay Body as a Bacterial Subcellular Structure marking multidrug tolerant Persisters

2018 ◽  
Author(s):  
Jiayu Yu ◽  
Yang Liu ◽  
Huijia Yin ◽  
Zengyi Chang
Keyword(s):  
Infectious Diseases ◽  
Bacterial Cells ◽  
Subcellular Structure ◽  
The Status ◽  
Multidrug Tolerance ◽  
Metabolic Activities ◽  
Accidental Discovery

SummaryBacteria have long been recognized to be capable of entering a phenotypically non-growing persister state, in which the cells exhibit an extended regrowth lag and a multidrug tolerance, thus posing a great challenge in treating infectious diseases. Owing to their non-inheritability, low abundance of existence, lack of metabolic activities, and high heterogeneity, properties of persisters remain poorly understood. Here, we report our accidental discovery of a hitherto unreported subcellular structure that we term the regrowth-delay body, which is formed only in non-growing bacterial cells and sequesters multiple key proteins. As of now, this structure, that dissolves when the cell resumes growth, is the most distinguishable subcellular structure marking persisters. Our studies also indicate that persisters exhibit different depth of persistence, as determined by the status of their regrowth-delay bodies. Our findings imply that suppressing the formation and/or promoting the dissolution of regrowth-delay bodies could be viable strategies for eradicating persisters.

Effect of piperacillin on morphology and viability of gram-negative bacteria

1984 ◽  
Vol 42 ◽  
pp. 218-219
Author(s):  
R. H. Liss
Keyword(s):  
Inhibitory Concentration ◽  
Cytoplasmic Membrane ◽  
Drug Binding ◽  
Gram Negative Bacteria ◽  
Bacterial Cells ◽  
Drug Induced ◽  
Penicillin Binding Proteins ◽  
Lactam Antibiotic ◽  
Drug Free ◽  
Tube Dilution

Piperacillip (PIP) is b-[D(-)-α-(4-ethy1-2,3-dioxo-l-piperzinylcar-bonylamino)-α-phenylacetamido]-penicillanate. The broad spectrum semisynthetic β-lactam antibiotic is believed to effect bactericidal activity through its affinity for penicillin-binding proteins (PBPs), enzymes on the bacterial cytoplasmic membrane that control elongation and septation during cell growth and division. The purpose of this study was to correlate penetration and binding of 14C-PIP in bacterial cells with drug-induced lethal changes assessed by microscopic, microbiologic and biochemical methods.The bacteria used were clinical isolates of Escherichia coli and Pseudomonas aeruginosa (Figure 1). Sensitivity to the drug was determined by serial tube dilution in Trypticase Soy Broth (BBL) at an inoculum of 104 organisms/ml; the minimum inhibitory concentration of piperacillin for both bacteria was 1 μg/ml. To assess drug binding to PBPs, the bacteria were incubated with 14C-PIP (5 μg/0.09 μCi/ml); controls, in drug-free medium.

Attachment of oral bacteria to titanium surfaces: An SEM study

1994 ◽  
Vol 52 ◽  
pp. 468-469
Author(s):  
J. E. Laffoon ◽  
R. L. Anderson ◽  
J. C. Keller ◽  
C. D. Wu-Yuan
Keyword(s):  
Technical Problem ◽  
Titanium Implant ◽  
Oral Bacteria ◽  
Bacterial Cells ◽  
Thin Sections ◽  
Surface Ultrastructure ◽  
Sem Study ◽  
Key Factor ◽  
Titanium Surfaces

Titanium (Ti) dental implants have been used widely for many years. Long term implant failures are related, in part, to the development of peri-implantitis frequently associated with bacteria. Bacterial adherence and colonization have been considered a key factor in the pathogenesis of many biomaterial based infections. Without the initial attachment of oral bacteria to Ti-implant surfaces, subsequent polymicrobial accumulation and colonization leading to peri-implant disease cannot occur. The overall goal of this study is to examine the implant-oral bacterial interfaces and gain a greater understanding of their attachment characteristics and mechanisms. Since the detailed cell surface ultrastructure involved in attachment is only discernible at the electron microscopy level, the study is complicated by the technical problem of obtaining titanium implant and attached bacterial cells in the same ultra-thin sections. In this study, a technique was developed to facilitate the study of Ti implant-bacteria interface.Discs of polymerized Spurr’s resin (12 mm x 5 mm) were formed to a thickness of approximately 3 mm using an EM block holder (Fig. 1). Titanium was then deposited by vacuum deposition to a film thickness of 300Å (Fig. 2).

Mechanisms involved in effects of antimicrobial peptides, bactenectins ChBac3.4, ChBac5, and mini-ChBac7.5Nb, on bacterial cells

2017 ◽  
pp. 43-50
Author(s):  
О.В. Шамова ◽  
М.С. Жаркова ◽  
П.М. Копейкин ◽  
Д.С. Орлов ◽  
Е.А. Корнева
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Activity ◽  
Innate Immunity ◽  
Infectious Diseases ◽  
Metabolic Activity ◽  
Capra Hircus ◽  
Bacterial Cells ◽  
Antibiotic Resistant ◽  
Resistant Strains

Антимикробные пептиды (АМП) системы врожденного иммунитета - соединения, играющие важную роль в патогенезе инфекционных заболеваний, так как обладают свойством инактивировать широкий спектр патогенных бактерий, обеспечивая противомикробную защиту живых организмов. В настоящее время АМП рассматриваются как потенциальные соединения-корректоры инфекционной патологии, вызываемой антибиотикорезистентными бактериями (АБР). Цель данной работы состояла в изученим механизмов антибактериального действия трех пептидов, принадлежащих к семейству бактенецинов - ChBac3.4, ChBac5 и mini-ChBac7.5Nb. Эти химически синтезированные пептиды являются аналогами природных пролин-богатых АМП, обнаруженных в лейкоцитах домашней козы Capra hircus и проявляющих высокую антимикробную активность, в том числе и в отношении грамотрицательных АБР. Методы. Минимальные ингибирующие и минимальные бактерицидные концентрации пептидов (МИК и МБК) определяли методом серийных разведений в жидкой питательной среде с последующим высевом на плотную питательную среду. Эффекты пептидов на проницаемость цитоплазматической мембраны бактерий для хромогенного маркера исследовали с использованием генетически модифицированного штамма Escherichia coli ML35p. Действие бактенецинов на метаболическую активность бактерий изучали с применением маркера резазурина. Результаты. Показано, что все исследованные пептиды проявляют высокую антимикробную активность в отношении Escherichia coli ML35p и антибиотикоустойчивых штаммов Escherichia coli ESBL и Acinetobacter baumannii in vitro, но их действие на бактериальные клетки разное. Использован комплекс методик, позволяющих наблюдать в режиме реального времени динамику действия бактенецинов в различных концентрациях (включая их МИК и МБК) на барьерную функцию цитоплазматической мембраны и на интенсивность метаболизма бактериальных клеток, что дало возможность выявить различия в характере воздействия бактенецинов, отличающихся по структуре молекулы, на исследуемые микроорганизмы. Установлено, что действие каждого из трех исследованных бактенецинов в бактерицидных концентрациях отличается по эффективности нарушения целостности бактериальных мембран и в скорости подавления метаболизма клеток. Заключение. Полученная информация дополнит существующие фундаментальные представления о механизмах действия пролин-богатых пептидов врожденного иммунитета, а также послужит основой для биотехнологических исследований, направленных на разработку на базе этих соединений новых антибиотических препаратов для коррекции инфекционных заболеваний, вызываемых АБР и являющимися причинами тяжелых внутрибольничных инфекций. Antimicrobial peptides (AMPs) of the innate immunity are compounds that play an important role in pathogenesis of infectious diseases due to their ability to inactivate a broad array of pathogenic bacteria, thereby providing anti-microbial host defense. AMPs are currently considered promising compounds for treatment of infectious diseases caused by antibiotic-resistant bacteria. The aim of this study was to investigate molecular mechanisms of the antibacterial action of three peptides from the bactenecin family, ChBac3.4, ChBac5, and mini-ChBac7.5Nb. These chemically synthesized peptides are analogues of natural proline-rich AMPs previously discovered by the authors of the present study in leukocytes of the domestic goat, Capra hircus. These peptides exhibit a high antimicrobial activity, in particular, against antibiotic-resistant gram-negative bacteria. Methods. Minimum inhibitory and minimum bactericidal concentrations of the peptides (MIC and MBC) were determined using the broth microdilution assay followed by subculturing on agar plates. Effects of the AMPs on bacterial cytoplasmic membrane permeability for a chromogenic marker were explored using a genetically modified strain, Escherichia coli ML35p. The effect of bactenecins on bacterial metabolic activity was studied using a resazurin marker. Results. All the studied peptides showed a high in vitro antimicrobial activity against Escherichia coli ML35p and antibiotic-resistant strains, Escherichia coli ESBL and Acinetobacter baumannii, but differed in features of their action on bacterial cells. The used combination of techniques allowed the real-time monitoring of effects of bactenecin at different concentrations (including their MIC and MBC) on the cell membrane barrier function and metabolic activity of bacteria. The differences in effects of these three structurally different bactenecins on the studied microorganisms implied that these peptides at bactericidal concentrations differed in their capability for disintegrating bacterial cell membranes and rate of inhibiting bacterial metabolism. Conclusion. The obtained information will supplement the existing basic concepts on mechanisms involved in effects of proline-rich peptides of the innate immunity. This information will also stimulate biotechnological research aimed at development of new antibiotics for treatment of infectious diseases, such as severe in-hospital infections, caused by antibiotic-resistant strains.

Process Description of an Unconventional Biofilm Formation by Bacterial Cells Autoagglutinating Through Sticky, Long, and Peritrichate Nanofibers

2019 ◽  
Author(s):  
Yoshihide Furuichi ◽  
Shogo Yoshimoto ◽  
Tomohiro Inaba ◽  
Nobuhiko Nomura ◽  
Katsutoshi Hori
Keyword(s):  
Formation Process ◽  
Biofilm Formation ◽  
Extracellular Polymeric Substances ◽  
Waste Treatment ◽  
Large Cell ◽  
Dlvo Theory ◽  
Bacterial Cells ◽  
Chemical Production ◽  
Discontinuous Surface ◽  
Cell Cell

<p></p><p>Biofilms are used in environmental biotechnologies including waste treatment and environmentally friendly chemical production. Understanding the mechanisms of biofilm formation is essential to control microbial behavior and improve environmental biotechnologies. <i>Acinetobacter </i>sp. Tol 5 autoagglutinate through the interaction of the long, peritrichate nanofiber protein AtaA, a trimeric autotransporter adhesin. Using AtaA, without cell growth or the production of extracellular polymeric substances, Tol 5 cells quickly form an unconventional biofilm. In this study, we investigated the formation process of this unconventional biofilm, which started with cell–cell interactions, proceeded to cell clumping, and led to the formation of large cell aggregates. The cell–cell interaction was described by DLVO theory based on a new concept, which considers two independent interactions between two cell bodies and between two AtaA fiber tips forming a virtual discontinuous surface. If cell bodies cannot collide owing to an energy barrier at low ionic strengths but approach within the interactive distance of AtaA fibers, cells can agglutinate through their contact. Cell clumping proceeds following the cluster–cluster aggregation model, and an unconventional biofilm containing void spaces and a fractal nature develops. Understanding its formation process would extend the utilization of various types of biofilms, enhancing environmental biotechnologies.</p><p></p>

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