METHODS FOR INDICATION OF NONCULTURATED VIABLE MICROORGANISMS WHEN CONTROL OF CRITICAL POINTS OF LIVESTOCK, POULTRY AND FOOD PRODUCTION TECHNOLOGY

2021 ◽  
Vol 1 (4) ◽  
pp. 441-447
Author(s):  
Ekaterina M. Lenchenko ◽  
◽  
Damir I. Udavliev ◽  
Inna B. Pavlova ◽  
◽  
...  
Keyword(s):  
Cell Wall ◽  
Luminescence Spectrum ◽  
Three Dimensional ◽  
Yeast Cells ◽  
Dimensional Structure ◽  
Heterogeneous Structure ◽  
Bacterial Cells ◽  
Microscopic Fungi ◽  
Gram Positive Bacteria ◽  
Red Luminescence

The results of morphometric and densitometric parameters biofilms are presented, effective methods of detecting uncultivated viable microorganisms isolated from a representative sample of objects of veterinary and sanitary supervision are tested and selected. Optical, luminescent and scanning electron microscopy revealed the formation of a three-dimensional structure biofilms in the form a dense network consisting of gram-negative and gram-positive bacteria, yeast cells, hyphal and pseudohyphalic forms, surrounded by an intercellular polymer matrix. The presence hyphae of microscopic fungi causes an increase in the number of cells adhered to the substrate, microcolonies were formed from bacteria and yeast cells of microscopic fungi. The pathogenesis of the syndrome of overgrowth of microorganisms is provided by the presence of various dissociative variants, the dispersion of uncultivated bacterial cells, which gain advantages in the hyperagregation of the architectonics of heterogeneous biofilms. Multilayer membranes, vesicles, cells with a defective cell wall, spheroplasts, protoplasts, L-shapes, needle-like and giant structures, and revertant cells were identified. The dynamics of changes in the viable structures microorganisms was characterized by alternating periods of decrease and increase in the intensity of biofilm formation. When detecting the viability of microorganisms in the composition biofilms, viable and non-viable cells were differentiated – a green luminescence spectrum and a red luminescence spectrum, respectively. The dissociation of the population caused an increase in the concentration of R-dissociant cells with a higher growth rate, cell lysis was detected after 48–72 h of cultivation, a change in the ratio phenotypic forms was observed – the M-dissociant was predominant. The study of the heterogeneous structure of the population, without disturbing the natural architectonics of biofilms, revealed direct correlations (r = 0,89) between morphometric (≥90 % of the field of view) and densitometric parameters (OD). The efficiency of a nutrient medium containing pancreatic hydrolyzate, mannitol, L-asparagine and glycerol was established for the repair of the cell wall, the reversal of L-forms of microorganisms.

scholarly journals Studies of biofilms and phenotypic characteristics of Candida fungi

2020 ◽  
pp. 132-138
Author(s):  
E. M. Lenchenko ◽  
N. P. Sachivkina
Keyword(s):  
Polymer Matrix ◽  
Polymer Matrix Composites ◽  
Three Dimensional ◽  
Significant Degree ◽  
Yeast Cells ◽  
Dimensional Structure ◽  
Heterogeneous Structure ◽  
Matrix Composites ◽  
Systemic Candidiasis ◽  
Candida Spp

Yeast-like fungi of the genus Candida are causative agents of the infectious pathology of the mucous membrane of the gastrointestinal, respiratory, urogenital tracts and skin of mammals, sepsis, and disseminated infection in birds. The search and testing of multilevel algorithms for biofilm identification when exposed to chemotherapeutic and disinfectant drugs for blocking the synthesis or destruction of the intercellular matrix in the development of superficial, deep and systemic candidiasis of animals are relevant for developing and improving diagnostic and antiepidemic measures. It was established that the formation of biofilm heterogeneous structure comprises multiple stages implementing the processes of intercellular communication due to the synthesis of a polymer matrix composites. Optical microscopy revealed a three-dimensional structure of biofilms in the form of a dense network consisting of yeast cells, hyphal and pseudohyphalic forms surrounded by an intercellular polymer matrix. Candida spp. pathogenicity factors contribute to infection of susceptible species due to adhesion, invasion, secretion of hydrolases, dimorphism. Formation of mono-species or poly-species biofilms of microorganisms, including Candida spp., causes the development of superficial, deep and systemic candidiasis. Detection of a large amount of yeast and micellar phases in C. albicans and C. africana isolates was a differential sign of a significant degree of colonization of the mucous membranes of the larynx, pharynx, and tonsils in case of local and systemic pathologies in pigs. The results of studies of the biofilm heterogeneous structure and phenotypic signs of yeast-like fungi can be used in a comparative study of biological characteristics and the identification of common patterns and differential signs of microorganisms, optimization of mycological diagnostics, and also in the development of antimycotic drugs.

scholarly journals Cryo-Electron Tomography Reveals the Comparative Three-Dimensional Architecture of Prochlorococcus, a Globally Important Marine Cyanobacterium

2007 ◽  
Vol 189 (12) ◽  
pp. 4485-4493 ◽  
Author(s):  
Claire S. Ting ◽  
Chyongere Hsieh ◽  
Sesh Sundararaman ◽  
Carmen Mannella ◽  
Michael Marko
Keyword(s):  
Cell Wall ◽  
Electron Tomography ◽  
Three Dimensional ◽  
Membrane System ◽  
Niche Differentiation ◽  
Dimensional Structure ◽  
Comparative Genomic ◽  
Photosynthetic Membrane ◽  
Peptidoglycan Biosynthesis ◽  
Cryo Electron Tomography

ABSTRACT In an age of comparative microbial genomics, knowledge of the near-native architecture of microorganisms is essential for achieving an integrative understanding of physiology and function. We characterized and compared the three-dimensional architecture of the ecologically important cyanobacterium Prochlorococcus in a near-native state using cryo-electron tomography and found that closely related strains have diverged substantially in cellular organization and structure. By visualizing native, hydrated structures within cells, we discovered that the MED4 strain, which possesses one of the smallest genomes (1.66 Mbp) of any known photosynthetic organism, has evolved a comparatively streamlined cellular architecture. This strain possesses a smaller cell volume, an attenuated cell wall, and less extensive intracytoplasmic (photosynthetic) membrane system compared to the more deeply branched MIT9313 strain. Comparative genomic analyses indicate that differences have evolved in key structural genes, including those encoding enzymes involved in cell wall peptidoglycan biosynthesis. Although both strains possess carboxysomes that are polygonal and cluster in the central cytoplasm, the carboxysomes of MED4 are smaller. A streamlined cellular structure could be advantageous to microorganisms thriving in the low-nutrient conditions characteristic of large regions of the open ocean and thus have consequences for ecological niche differentiation. Through cryo-electron tomography we visualized, for the first time, the three-dimensional structure of the extensive network of photosynthetic lamellae within Prochlorococcus and the potential pathways for intracellular and intermembrane movement of molecules. Comparative information on the near-native structure of microorganisms is an important and necessary component of exploring microbial diversity and understanding its consequences for function and ecology.

scholarly journals Three-dimensional structure of the bacterial cell wall peptidoglycan

2006 ◽  
Vol 103 (12) ◽  
pp. 4404-4409 ◽  
Author(s):  
S. O. Meroueh ◽  
K. Z. Bencze ◽  
D. Hesek ◽  
M. Lee ◽  
J. F. Fisher ◽  
...  
Keyword(s):  
Cell Wall ◽  
Bacterial Cell ◽  
Three Dimensional ◽  
Bacterial Cell Wall ◽  
Dimensional Structure ◽  
Three Dimensional Structure ◽  
Cell Wall Peptidoglycan

scholarly journals Crystallization and preliminary crystallographic analysis of human muscle phosphofructokinase, the main regulator of glycolysis

2014 ◽  
Vol 70 (5) ◽  
pp. 578-582 ◽  
Author(s):  
Marco Kloos ◽  
Antje Brüser ◽  
Jürgen Kirchberger ◽  
Torsten Schöneberg ◽  
Norbert Sträter
Keyword(s):  
Three Dimensional ◽  
Crystallographic Analysis ◽  
Human Muscle ◽  
Yeast Cells ◽  
Dimensional Structure ◽  
Structural Basis ◽  
Diffusion Method ◽  
Rabbit Muscle ◽  
Main Regulator ◽  
Purification Protocol

Whereas the three-dimensional structure and the structural basis of the allosteric regulation of prokaryotic 6-phosphofructokinases (Pfks) have been studied in great detail, knowledge of the molecular basis of the allosteric behaviour of the far more complex mammalian Pfks is still very limited. The human muscle isozyme was expressed heterologously in yeast cells and purified using a five-step purification protocol. Protein crystals suitable for diffraction experiments were obtained by the vapour-diffusion method. The crystals belonged to space groupP6222 and diffracted to 6.0 Å resolution. The 3.2 Å resolution structure of rabbit muscle Pfk (rmPfk) was placed into the asymmetric unit and optimized by rigid-body and groupB-factor refinement. Interestingly, the tetrameric enzyme dissociated into a dimer, similar to the situation observed in the structure of rmPfk.

scholarly journals Bacterial envelope damage inflicted by bioinspired nanospikes grown in a hydrogel

2020 ◽  
Author(s):  
Sandra L. Arias ◽  
Joshua Devorkin ◽  
Jessica C. Spear ◽  
Ana Civantos ◽  
Jean Paul Allain
Keyword(s):  
Mechanical Properties ◽  
Antimicrobial Activity ◽  
Cell Wall ◽  
Mammalian Cells ◽  
Cell Shape ◽  
Rational Design ◽  
Bacterial Cells ◽  
Nanoscale Structures ◽  
Gram Positive Bacteria ◽  
Tissue Restoration

AbstractDevice-associated infections are one of the deadliest complications accompanying the use of biomaterials, and despite recent advances in the development of anti-biofouling strategies, biomaterials that exhibit both functional tissue restoration and antimicrobial activity have been challenging to achieve. Here, we report the fabrication of bio-inspired bactericidal nanospikes in bacterial cellulose and investigate the mechanism underlying this phenomenon. We demonstrate these structures affects preferentially stiff membranes like those in Gram-positive bacteria, but exhibit cytocompatibility towards mammalian cells, a requisite for tissue restoration. We also reveal the bactericidal activity of the nanospikes is due to a pressure-induced mechanism, which depends on the cell’s adherence time, nanospike’s geometry and spacing, cell shape, and mechanical properties of the cell wall. Our findings provide a better understanding of the mechanobiology of bacterial cells at the interface with nanoscale structures, which is fundamental for the rational design bactericidal topographies.

scholarly journals Spatial mapping of immunological epitopes in the Candida cell wall using C-type lectin probes

2021 ◽  
Vol 3 (12) ◽  
Author(s):  
Ingrida Vendele ◽  
Ten Feizi ◽  
Maria Spyrou ◽  
Mark Stappers ◽  
Gordon Brown ◽  
...  
Keyword(s):  
Cell Wall ◽  
Three Dimensional ◽  
Fungal Species ◽  
Physiological Adaptation ◽  
Dimensional Structure ◽  
Immune Reactivity ◽  
Fungal Cell ◽  
Lectin Receptor ◽  
Composition Changes

The primary recognition event between a fungal pathogen and the immune system normally involves the engagement of a pattern recognition receptor with specific components of the cell wall. However, the cell wall is a complex three dimensional structure whose composition changes rapidly in accordance with environmental stimuli. Therefore it is important to know what is the precise nature of the primary recognition event, how many events occur to activate the immune response and how these recognition events are affected by changes in cell wall architecture, cellular morphogenesis and physiological adaptation of the pathogen to specific niches in the human body. We address this fundamental question using four soluble immune C-Type lectin receptor-probes which recognize specific mannans and β-1,3 glucan in the cell wall. We use these C-type lectin probes to demonstrate that mannan epitopes are differentially distributed in the inner and outer layers of fungal cell wall in a clustered or diffuse manner. Immune reactivity of fungal cell surfaces did not correlate with relatedness of different fungal species, and mannan-detecting receptor-probes discriminated between cell surface mannans generated by the same fungus growing under different conditions. These studies demonstrate that mannan-epitopes within fungal cell walls are differentially distributed and dynamically expressed as the fungus adapted to microenvironments that would be encountered in vivo.

The three-dimensional structure of the cell wall glycoprotein of Chlorogonium elongatum

1984 ◽  
Vol 68 (1) ◽  
pp. 271-284
Author(s):  
P.J. Shaw ◽  
G.J. Hills
Keyword(s):  
Cell Wall ◽  
Plant Cell Wall ◽  
Higher Plants ◽  
Three Dimensional ◽  
Three Dimensions ◽  
Dimensional Structure ◽  
Primary Cell Wall ◽  
Globular Domain ◽  
Higher Plant ◽  
Intersubunit Interactions

The green alga Chlorogonium elongatum, a member of the Volvocales, possesses a crystalline cell wall composed of hydroxyproline-rich glycoprotein similar to the primary cell wall glycoproteins of higher plants. Electron microscopy and computer image processing have been used to determine the crystal structure of the Chlorogonium cell wall in three dimensions to a resolution of 2.0 nm. The structure is composed of heterologous dimers. Each subunit of the dimer comprises a long, thin spacer domain and a large globular domain, which is the site of the intra- and inter-dimer interactions. There are also sites of intersubunit interactions at the opposite ends of the rod domains. We suggest that the rods are composed predominantly of glycosylated polyproline helix, as has been suggested for higher plant cell wall glycoproteins and has been shown for the cell wall glycoprotein of Chlamydomonas reinhardtii, which is closely related to Chlorogonium.

scholarly journals The clustering of telomeres and colocalization with Rap1, Sir3, and Sir4 proteins in wild-type Saccharomyces cerevisiae.

1996 ◽  
Vol 134 (6) ◽  
pp. 1349-1363 ◽  
Author(s):  
M Gotta ◽  
T Laroche ◽  
A Formenton ◽  
L Maillet ◽  
H Scherthan ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequences ◽  
Three Dimensional ◽  
Yeast Cells ◽  
Dimensional Structure ◽  
Nuclear Pore ◽  
Wild Type ◽  
Telomeric Silencing ◽  
Sites Of Action

We have developed a novel technique for combined immunofluorescence/in situ hybridization on fixed budding yeast cells that maintains the three-dimensional structure of the nucleus as monitored by focal sections of cells labeled with fluorescent probes and by staining with a nuclear pore antibody. Within the resolution of these immunodetection techniques, we show that proteins encoded by the SIR3, SIR4, and RAP1 genes colocalize in a statistically significant manner with Y' telomere-associated DNA sequences. In wild-type cells the Y' in situ hybridization signals can be resolved by light microscopy into fewer than ten foci per diploid nucleus. This suggests that telomeres are clustered in vegetatively growing cells, and that proteins essential for telomeric silencing are concentrated at their sites of action, i.e., at telomeres and/or subtelomeric regions. As observed for Rap1, the Sir4p staining is diffuse in a sir3- strain, and similarly, Sir3p staining is no longer punctate in a sir4- strain, although the derivatized Y' probe continues to label discrete sites in these strains. Nonetheless, the Y' FISH is altered in a qualitative manner in sir3 and sir4 mutant strains, consistent with the previously reported phenotypes of shortened telomeric repeats and loss of telomeric silencing.

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