scholarly journals Characterization of Mono- and Mixed-Culture Campylobacter jejuni Biofilms

2011 ◽  
Vol 78 (4) ◽  
pp. 1033-1038 ◽  
Author(s):  
Tuba Ica ◽  
Vildan Caner ◽  
Ozlem Istanbullu ◽  
Hung Duc Nguyen ◽  
Bulbul Ahmed ◽  
...  

ABSTRACTCampylobacter jejuni, one of the most common causes of human gastroenteritis, is a thermophilic and microaerophilic bacterium. These characteristics make it a fastidious organism, which limits its ability to survive outside animal hosts. Nevertheless,C. jejunican be transmitted to both humans and animals via environmental pathways, especially through contaminated water. Biofilms may play a crucial role in the survival of the bacterium under unfavorable environmental conditions. The goal of this study was to investigate survival strategies ofC. jejuniin mono- and mixed-culture biofilms. We grew monoculture biofilms ofC. jejuniand mixed-culture biofilms ofC. jejuniwithPseudomonas aeruginosa. We found that mono- and mixed-culture biofilms had significantly different structures and activities. MonocultureC. jejunibiofilms did not consume a measurable quantity of oxygen. Using a confocal laser scanning microscope (CLSM), we found that cells from monoculture biofilms were alive according to live/dead staining but that these cells were not culturable. In contrast, in mixed-culture biofilms,C. jejuniremained in a culturable physiological state. MonocultureC. jejunibiofilms could persist under lower flow rates (0.75 ml/min) but were unable to persist at higher flow rates (1 to 2.5 ml/min). In sharp contrast, mixed-culture biofilms were more robust and were unaffected by higher flow rates (2.5 ml/min). Our results indicate that biofilms provide an environmental refuge that is conducive to the survival ofC. jejuni.

2021 ◽  
Vol ahead-of-print (ahead-of-print) ◽  
Author(s):  
Zhong Li ◽  
Qing Lei ◽  
Luyao Huang ◽  
Chao Liu

Purpose Low-alloy structural steels (LASS) face severe microbiologically influenced corrosion (MIC) in their service environments. To mitigate this issue, Cu is often used as an alloying element owing to its intrinsic antimicrobial activity. However, the antibacterial performance and biofilm resistance of Cu-containing LASS (Cu-LASS) are still unclear. This study aims to analyze the effect of Cu addition to 420 MP LASS on its MIC by the Pseudomonas aeruginosa biofilm. Design/methodology/approach Scanning electron microscope, confocal laser scanning microscope and X-ray photoelectron spectroscopy were used to analyze the surface morphology and composition of corrosion products. The antibacterial activities of Cu-LASS were analyzed by the spread-plate method. In addition, electrochemical analysis was conducted to characterize the corrosion behavior of the produced alloy. Findings Bacterial analysis and morphological observation confirmed a reduced sessile cell count and inactivation of the P. aeruginosa biofilm on the surface of Cu-LASS coupons. Electrochemical measurements showed that Cu-LASS exhibited large polarization and charge-transfer resistances, which indicated excellent MIC resistance. This significantly enhanced resistance to MIC could be explained by the synergistic effect of released Cu2+ from the Cu-LASS surface and immediate contact to Cu-rich phase in the surface and the release of Cu2+ ions from the Cu-LASS surface. Originality/value The effect of Cu addition on the MIC resistance and antibacterial performance of LASS is seldom reported. It is necessary to investigate the corrosion resistance of Cu-LASS and clarify its antibacterial mechanism. This paper fulfills this need.


2011 ◽  
Vol 78 (1) ◽  
pp. 7-13 ◽  
Author(s):  
E. L. Ramírez-Reinat ◽  
F. Garcia-Pichel

ABSTRACTRecent physiological work has shown that the filamentous euendolithic cyanobacteriumMastigocoleus testarum(strain BC008) is able to bore into solid carbonates using Ca2+-ATPases to take up Ca2+from the medium at the excavation front, promoting dissolution of CaCO3there. It is not known, however, if this is a widespread mechanism or, rather, a unique capability of this model strain. To test this, we undertook a survey of multispecies euendolithic microbial assemblages infesting natural carbonate substrates in marine coastal waters of the Caribbean, Mediterranean, South Pacific, and Sea of Cortez. Microscopic examination revealed the presence of complex assemblages of euendoliths, encompassing 3 out of the 5 major cyanobacterial orders. 16S rRNA gene clone libraries detected even greater diversity, particularly among the thin-filamentous forms, and allowed us to categorize the endoliths in our samples into 8 distinct phylogenetic clades. Using real-time Ca2+imaging under a confocal laser scanning microscope, we could show that all communities displayed light-dependent formation of Ca2+-supersaturated zones in and around boreholes, a staple of actively boring phototrophs. In 3 out of 4 samples, boring activity was sensitive to at least one of two inhibitors of Ca2+-ATPase transporters (thapsigargin ortert-butylhydroquinone), indicating that the Ca2+-ATPase mechanism is widespread among cyanobacterial euendoliths but perhaps not universal. Function-community structure correlations point to one particular clade of baeocyte-forming euendoliths as the potential exception.


2013 ◽  
Vol 16 (2) ◽  
pp. 387-389
Author(s):  
A. Sałamaszyńska-Guz ◽  
M.M. Godlewski ◽  
D. Klimuszko

AbstractThe cj0183 and cj0588 genes identified in the Campylobacter jejuni NCTC 11168 genome encode proteins with homology to virulence factors found in other bacteria. Previous studies showed that single mutation in the cj0183 gene does not affect adhesion of C. jejuni to the Caco-2 cell line whereas protein encoded by cj0588 is involved in adherence to the Caco-2 cells. In the presented study differences in invasion index were observed between mutants in both genes and single mutation of cj0588 in 81116 and 81-176 C. jejuni strains. This fact indicates that Cj0183 protein might play some role in invasion of bacteria into host cells.


Author(s):  
Thomas M. Jovin ◽  
Michel Robert-Nicoud ◽  
Donna J. Arndt-Jovin ◽  
Thorsten Schormann

Light microscopic techniques for visualizing biomolecules and biochemical processes in situ have become indispensable in studies concerning the structural organization of supramolecular assemblies in cells and of processes during the cell cycle, transformation, differentiation, and development. Confocal laser scanning microscopy offers a number of advantages for the in situ localization and quantitation of fluorescence labeled targets and probes: (i) rejection of interfering signals emanating from out-of-focus and adjacent structures, allowing the “optical sectioning” of the specimen and 3-D reconstruction without time consuming deconvolution; (ii) increased spatial resolution; (iii) electronic control of contrast and magnification; (iv) simultanous imaging of the specimen by optical phenomena based on incident, scattered, emitted, and transmitted light; and (v) simultanous use of different fluorescent probes and types of detectors.We currently use a confocal laser scanning microscope CLSM (Zeiss, Oberkochen) equipped with 3-laser excitation (u.v - visible) and confocal optics in the fluorescence mode, as well as a computer-controlled X-Y-Z scanning stage with 0.1 μ resolution.


Author(s):  
Thomas J. Deerinck ◽  
Maryann E. Martone ◽  
Varda Lev-Ram ◽  
David P. L. Green ◽  
Roger Y. Tsien ◽  
...  

The confocal laser scanning microscope has become a powerful tool in the study of the 3-dimensional distribution of proteins and specific nucleic acid sequences in cells and tissues. This is also proving to be true for a new generation of high contrast intermediate voltage electron microscopes (IVEM). Until recently, the number of labeling techniques that could be employed to allow examination of the same sample with both confocal and IVEM was rather limited. One method that can be used to take full advantage of these two technologies is fluorescence photooxidation. Specimens are labeled by a fluorescent dye and viewed with confocal microscopy followed by fluorescence photooxidation of diaminobenzidine (DAB). In this technique, a fluorescent dye is used to photooxidize DAB into an osmiophilic reaction product that can be subsequently visualized with the electron microscope. The precise reaction mechanism by which the photooxidation occurs is not known but evidence suggests that the radiationless transfer of energy from the excited-state dye molecule undergoing the phenomenon of intersystem crossing leads to the formation of reactive oxygen species such as singlet oxygen. It is this reactive oxygen that is likely crucial in the photooxidation of DAB.


Author(s):  
P.M. Houpt ◽  
A. Draaijer

In confocal microscopy, the object is scanned by the coinciding focal points (confocal) of a point light source and a point detector both focused on a certain plane in the object. Only light coming from the focal point is detected and, even more important, out-of-focus light is rejected.This makes it possible to slice up optically the ‘volume of interest’ in the object by moving it axially while scanning the focused point light source (X-Y) laterally. The successive confocal sections can be stored in a computer and used to reconstruct the object in a 3D image display.The instrument described is able to scan the object laterally with an Ar ion laser (488 nm) at video rates. The image of one confocal section of an object can be displayed within 40 milliseconds (1000 х 1000 pixels). The time to record the total information within the ‘volume of interest’ normally depends on the number of slices needed to cover it, but rarely exceeds a few seconds.


TAPPI Journal ◽  
2010 ◽  
Vol 9 (10) ◽  
pp. 7-15
Author(s):  
HANNA KOIVULA ◽  
DOUGLAS BOUSFIELD ◽  
MARTTI TOIVAKKA

In the offset printing process, ink film splitting has an important impact on formation of ink filaments. The filament size and its distribution influence the leveling of ink and hence affect ink setting and the print quality. However, ink filaments are difficult to image due to their short lifetime and fine length scale. Due to this difficulty, limited work has been reported on the parameters that influence filament size and methods to characterize it. We imaged ink filament remains and quantified some of their characteristics by changing printing speed, ink amount, and fountain solution type. Printed samples were prepared using a laboratory printability tester with varying ink levels and operating settings. Rhodamine B dye was incorporated into fountain solutions to aid in the detection of the filaments. The prints were then imaged with a confocal laser scanning microscope (CLSM) and images were further analyzed for their surface topography. Modeling of the pressure pulses in the printing nip was included to better understand the mechanism of filament formation and the origin of filament length scale. Printing speed and ink amount changed the size distribution of the observed filament remains. There was no significant difference between fountain solutions with or without isopropyl alcohol on the observed patterns of the filament remains.


2019 ◽  
Vol 10 ◽  
pp. 2182-2191 ◽  
Author(s):  
Tushar C Jagadale ◽  
Dhanya S Murali ◽  
Shi-Wei Chu

Nonlinear nanoplasmonics is a largely unexplored research area that paves the way for many exciting applications, such as nanolasers, nanoantennas, and nanomodulators. In the field of nonlinear nanoplasmonics, it is highly desirable to characterize the nonlinearity of the optical absorption and scattering of single nanostructures. Currently, the common method to quantify optical nonlinearity is the z-scan technique, which yields real and imaginary parts of the permittivity by moving a thin sample with a laser beam. However, z-scan typically works with thin films, and thus acquires nonlinear responses from ensembles of nanostructures, not from single ones. In this work, we present an x-scan technique that is based on a confocal laser scanning microscope equipped with forward and backward detectors. The two-channel detection offers the simultaneous quantification for the nonlinear behavior of scattering, absorption and total attenuation by a single nanostructure. At low excitation intensities, both scattering and absorption responses are linear, thus confirming the linearity of the detection system. At high excitation intensities, we found that the nonlinear response can be derived directly from the point spread function of the x-scan images. Exceptionally large nonlinearities of both scattering and absorption are unraveled simultaneously for the first time. The present study not only provides a novel method for characterizing nonlinearity of a single nanostructure, but also reports surprisingly large plasmonic nonlinearities.


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