Gradients in the taxonomic composition of different microbial systems: comparison between biofilms for advanced waste treatment and lake sediments

1998 ◽  
Vol 37 (4-5) ◽  
pp. 159-166 ◽  
Author(s):  
I. Röske ◽  
K. Röske ◽  
D. Uhlmann
Keyword(s):  
Laser Scanning ◽  
Flow Reactor ◽  
Treatment Plant ◽  
Water Reservoir ◽  
Laser Scanning Microscopy ◽  
Microbial Colonization ◽  
Epifluorescence Microscopy ◽  
Confocal Laser ◽  
Sediment Layer ◽  
Glass Slides

The application of in situ hybridization with group specific oligonucleotide probes detected by epifluorescence microscopy and confocal laser scanning microscopy was tested to identify spatial gradients in the distribution of bacteria in biofilms of plug flow reactors and in the bottom sediment layer of a drinking water reservoir. The two tubular biofilm reactors were fed with the effluent from a full scale biological wastewater treatment plant to which were added the chlorophenols whole degradation was being investigated. One was operated as a continuous-flow reactor and the other as a sequencing batch reactor. The vertical gradients in the microbial colonization of the sediment were analyzed by means of glass slides exposed to the sediment. In the biofilms of both reactors the beta-Proteobacteria dominated. The Cytophaga-Flavobacterium group and the Gram-positive bacteria were also abundant. Only small amounts of gamma-bacteria could be detected. This is contrary to findings using traditional cultivation methods. Unlike the biofilms in the reactor, the bacterial Aufwuchs on the glass slides in the sediment presented a surprising diversity of morphological types and size classes of bacteria.

Culture and Detection of Campylobacter jejuni within Mixed Microbial Populations of Biofilms on Stainless Steel

2007 ◽  
Vol 70 (6) ◽  
pp. 1379-1385 ◽  
Author(s):  
SHERIASE Q. SANDERS ◽  
DOROTHY H. BOOTHE ◽  
JOSEPH F. FRANK ◽  
JUDY W. ARNOLD
Keyword(s):  
Stainless Steel ◽  
Campylobacter Jejuni ◽  
Laser Scanning ◽  
The United States ◽  
Laser Scanning Microscopy ◽  
Epifluorescence Microscopy ◽  
Confocal Laser ◽  
Atmospheric Conditions ◽  
Culture Methods ◽  
Bacterial Populations

Campylobacter jejuni is the most frequently reported cause of foodborne illness in the United States, but its survival outside the host is poor. The objective of this research was to examine the formation and composition of biofilms by C. jejuni alone and within mixed bacterial populations from the poultry-processing environment. C. jejuni growth was assessed with four media, two temperatures, and two atmospheric conditions to develop culture methods for liquid media that would allow growth within the biofilms. Growth kinetics was followed at four cell densities to determine temporal compatibility within biofilm mixtures. Analysis of the biofilms by confocal laser scanning microscopy showed that C. jejuni formed a biofilm when incubated without other bacteria. The average surface area of stainless steel covered by C. jejuni increased by 50% from 24 to 48 h, remained level to 96 h, and then decreased by 88% by 168 h. C. jejuni and mixed bacterial populations formed biofilms during incubation periods of up to 7 days. The area of the mixture was significantly greater than for C. jejuni alone at 24 h, was approximately the same at 48 h, and was significantly less by 168 h. When incubated with either of two initial inoculum densities of other bacteria, the number of C. jejuni was enhanced after 24 h. The intensity of fluorescence and cell viability were monitored by epifluorescence microscopy. This study provides the basis for studying interactions of Campylobacter spp. with other bacteria in the environment, which will aid in the design of effective intervention strategies.

Population dynamics of rumen microbes using modern techniques in rumen enhanced solid incubation

2003 ◽  
Vol 48 (4) ◽  
pp. 113-119 ◽  
Author(s):  
N. Raizada ◽  
V. Sonakya ◽  
R. Dalhoff ◽  
M. Hausner ◽  
P.A. Wilderer
Keyword(s):  
Population Dynamics ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Epifluorescence Microscopy ◽  
Rumen Content ◽  
Confocal Laser ◽  
Rumen Microbes ◽  
Microbial Population Dynamics ◽  
The Stability

The microbial ecology of the rumen is very complex. Different species of bacteria, protozoa, and fungi are involved in digestion of plant material in ruminants. In spite of complicated interrelationships among the various groups of microorganisms in the rumen ecosystem, Bacteria and Archaea are believed to play a major role because of their numerical predominance and metabolic diversity. In this work we are presenting the results for microbial population dynamics of rumen microbes during two-stage anaerobic digestion of grass. The reactors were inoculated with fresh rumen content. Fluorescent in situ hybridization, confocal laser scanning microscopy and epifluorescence microscopy were employed for microbial investigation. It was observed that Bacteria dominated in the hydrolytic reactor (1st stage) whereas Archaea were predominant in the methanogenic reactor (2nd stage). The stability of the methanogenic reactor was result of the dominance of Methanosaeta species (mainly the filamentous type).

scholarly journals Characterization and Quenching of Autofluorescence in Piglet Testis Tissue and Cells

2012 ◽  
Vol 2012 ◽  
pp. 1-10 ◽  
Author(s):  
Yanfei Yang ◽  
Ali Honaramooz
Keyword(s):  
Laser Scanning ◽  
Intrinsic Fluorescence ◽  
Laser Scanning Microscopy ◽  
Epifluorescence Microscopy ◽  
Fluorescence Labeling ◽  
Confocal Laser ◽  
Target Cells ◽  
Tissue Sections ◽  
Sudan Black B ◽  
Testis Tissue

Significant intrinsic fluorescence in tissues and in disassociated cells can interfere with fluorescence identification of target cells. The objectives of the present study were (1) to examine an intrinsic fluorescence we observed in both the piglet testis tissue and cells and (2) to test an effective method to block the autofluorescence. We observed that a number of granules within the testis interstitial cells were inherently fluorescent, detectable using epifluorescence microscopy, confocal laser scanning microscopy, and flow cytometry. The emission wavelength of the autofluorescent substance ranged from 425 to 700 nm, a range sufficiently broad that could potentially interfere with fluorescence techniques. When we treated the samples with Sudan Black B for different incubation times, the intrinsic fluorescence was completely masked after treatment for 10–15 min of the testis tissue sections or for 8 min of the testis cells, without compromising specific fluorescence labeling of gonocytes with lectin Dolichos biflorus agglutinin (DBA). We speculate that the lipofuscin or lipofuscin-like pigments within Leydig cell granules were mainly responsible for the observed intrinsic fluorescence in piglet testes. The method described in the present study can facilitate the identification and characterization of piglet gonocytes using fluorescence microscopy.

Attachment to Minerals and Biofilm Development of Extremely Acidophilic Archaea

2013 ◽  
Vol 825 ◽  
pp. 103-106 ◽  
Author(s):  
Rui Yong Zhang ◽  
Mario Vera ◽  
Sören Bellenberg ◽  
Wolfgang Sand
Keyword(s):  
Laser Scanning ◽  
Preferential Attachment ◽  
Growth Conditions ◽  
Laser Scanning Microscopy ◽  
Epifluorescence Microscopy ◽  
Confocal Laser ◽  
Planktonic Cells ◽  
Force Microscopy ◽  
Morphological Differences ◽  
Biofilm Development

Biofilm development of F. acidiphilum BRGM4 on polycarbonate filters floating on liquid medium and pyrite surfaces were studied by confocal laser scanning microscopy (CLSM) and atomic force microscopy (AFM) combined with epifluorescence microscopy (EFM). Results show that F. acidiphilum biofilms were heterogeneously distributed, and varied among different growth conditions, such as inorganic phosphate (Pi) starvation and glucose supplementation. Biofilm and planktonic cells showed significant morphological differences. Capsular EPS were observed in both biofilm and planktonic cells. Cells showed preferential attachment to the cracks/defects of pyrite surfaces.

Differential microbial colonization on microplastic in the Mediterranean Sea coastal zone

2020 ◽  
Author(s):  
Annika Vaksmaa ◽  
Katrin Knittel ◽  
Alejandro Abdala Asbun ◽  
Maaike Goudriaan ◽  
Andreas Ellrott ◽  
...  
Keyword(s):  
Mediterranean Sea ◽  
Laser Scanning ◽  
Mesh Size ◽  
Laser Scanning Microscopy ◽  
Microbial Colonization ◽  
Confocal Laser ◽  
Rrna Gene ◽  
Plastic Debris ◽  
Significant Difference ◽  
The Mediterranean

<p>Ocean plastic debris poses a large threat to the marine environment. Millions of tons of plastic end up in the ocean each year and the Mediterranean Sea is one of the most plastic polluted sea. Ocean plastic particles are typically covered with microbial biofilms, but it remains unclear if different polymer types are colonized by different communities. Knowledge in this aspect strengthens our understanding if microbes purely use plastic debris as attachment surface or if they may even contribute to the degradation of plastic. To gain a better understanding of the composition and structure of biofilms on micro plastic particles (MP) in the Mediterranean Sea, we analyzed microbial community covering floating MP in a bay/marina (Marina di Campo) on the island of Elba. MPs were collected with a plankton net (mesh size 50µm), fixed for fluorescence microscopy and stored for subsequent DNA extraction, and identification of the polymer with Raman spectroscopy. The particles were mainly comprised of polyethylene (PE), polypropylene (PP) and polystyrene (PS) and were often brittle and with cracks (PE, PP) and showed visual signs of biofouling (PE, PP, PS). Fluorescence in situ hybridization and imaging by high resolution confocal laser scanning microscopy of single MPs revealed high densities of colonization by microbes. 16S rRNA gene amplicon sequencing (Illumina Miseq) revealed higher abundance of archaeal sequences on PS (up to 29% of the reads) in comparison to PE or PP (up to 3% of the reads).  The bacterial community in the biofilms on each of the three plastic types consisted mainly of the orders Flavobacteriales, Rickettsiales, Alteromonadales, Cytophagales, Rhodobacterales and Oceanospirillales. Furthermore, we found significant difference in the community composition of biofilms on PE compared to PP and PS but not between PP and PS. The indicator species on PE were Calditrichales, detected at 10 times higher sequence abundance on PE than on PP and PS, as well as several uncultured orders. This study sheds light on preferential microbial attachment and biofilm formation on microplastic particles, yet it remains to be revealed, whether and which of these may contribute to plastic degradation.</p>

Stimulus-dependent relocation of the microtubule organizing center in human polymorphonuclear leukocytes

1992 ◽  
Vol 102 (4) ◽  
pp. 723-728
Author(s):  
J.M. Chiplonkar ◽  
D.D. Vandre ◽  
J.M. Robinson
Keyword(s):  
Polymorphonuclear Leukocytes ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Complex Iii ◽  
Epifluorescence Microscopy ◽  
Confocal Laser ◽  
Microtubule Organizing Center ◽  
Organizing Center ◽  
Antigen Antibody

Polymorphonuclear leukocytes (PMNs) exhibit extensive directional migration (chemotaxis) and phagocytic activities. We have developed an in vitro model to evaluate the organization of the microtubule organizing center (MTOC) in PMNs as the latter interact with various substrata, including immobilized antigen-antibody complexes. PMNs were layered on poly-L-lysine substrata containing ferritin (PL+F) or ferritin-antiferritin complex (PL+F+AF) and the location of MTOCs was determined by indirect immunofluorescence of tubulin using conventional epifluorescence microscopy and confocal laser scanning microscopy. The MTOCs in the majority of the PMNs attached to PL+F occupied an apical location (81.29% +/− 3.34%), while in the majority of PMNs layered onto PL+F+AF, a basal location (79.37% +/− 5.26%) was observed. Following disruption of microtubules (MTs) by nocodazole before layering the cells on the substrata, the proportions of PMNs with apical MTOCs were 65.2% +/− 6.27% for PL+F and 47.2% +/− 4.1% for PL+F+AF substrata, while the proportions of PMNs with basal MTOCs were 26.11% +/− 8.89% for PL+F and 39.6% +/− 4.4 for PL+F+AF substrata. The results indicate that MTOCs in human PMNs in vitro (i) occupied a ‘pre-defined’ apical location; (ii) translocated to a ‘newly defined’ basal location upon stimulation with immobilized antigen-antibody complex; (iii) and depended on intact MTs for placement of MTOCs in both situations.

scholarly journals Spatiotemporal Colonization of Xylella fastidiosa in its Vector Supports the Role of Egestion in the Inoculation Mechanism of Foregut-Borne Plant Pathogens

2011 ◽  
Vol 101 (8) ◽  
pp. 912-922 ◽  
Author(s):  
Elaine A. Backus ◽  
David J. W. Morgan
Keyword(s):  
Laser Scanning ◽  
Plant Pathogens ◽  
Fluorescent Protein ◽  
Xylella Fastidiosa ◽  
Functional Anatomy ◽  
Laser Scanning Microscopy ◽  
Microbial Colonization ◽  
Confocal Laser ◽  
Spatiotemporal Pattern ◽  
Homalodisca Vitripennis

The pathogen that causes Pierce's disease of grapevine, Xylella fastidiosa, is the only known bacterial, arthropod-transmitted plant pathogen that does not circulate in the vector's hemolymph. Instead, bacteria are foregut-borne, persistent in adult vectors but semipersistent in immatures (i.e., bacteria colonize cuticular surfaces of the anterior foregut, are retained for hours to days, but are lost during molting). Yet, exactly how a sharpshooter vector inoculates bacteria from foregut acquisition sites is unknown. The present study used confocal laser-scanning microscopy to identify locations in undissected, anterior foreguts of the glassy-winged sharpshooter colonized by green fluorescent protein-expressing X. fastidiosa. Spatial and temporal distributions of colonizing X. fastidiosa were examined daily over acquisition access periods of 1 to 6 days for both contaminated field-collected and clean laboratory-reared Homalodisca vitripennis. Results provide the first direct, empirical evidence that established populations of X. fastidiosa can disappear from vector foreguts over time. When combined with existing knowledge on behavior, physiology, and functional anatomy of sharpshooter feeding, present results support the idea that the disappearance is caused by outward fluid flow (egestion) not inward flow (ingestion) (i.e., swallowing). Thus, results support the hypothesis that egestion is a critical part of the X. fastidiosa inoculation mechanism. Furthermore, results suggest a cyclical, spatiotemporal pattern of microbial colonization, disappearance, and recolonization in the precibarium. Colonization patterns also support two types of egestion, termed rinsing and discharging egestion herein. Finally, comparison of acquisition results for field-collected versus laboratory-reared sharpshooters suggest that there may be competitive binding for optimum acquisition sites in the foregut. Therefore, successful inoculation of X. fastidiosa may depend, in large part, on vector load in the precibarium.

scholarly journals Comparison of the Antimicrobial Effects of Chlorine, Silver Ion, and Tobramycin on Biofilm

2008 ◽  
Vol 52 (4) ◽  
pp. 1446-1453 ◽  
Author(s):  
Jaeeun Kim ◽  
Betsey Pitts ◽  
Philip S. Stewart ◽  
Anne Camper ◽  
Jeyong Yoon
Keyword(s):  
Laser Scanning ◽  
Antimicrobial Agents ◽  
Respiratory Activity ◽  
Membrane Integrity ◽  
Laser Scanning Microscopy ◽  
Epifluorescence Microscopy ◽  
Confocal Laser ◽  
Tetrazolium Chloride ◽  
Antimicrobial Efficiency ◽  
Sequential Treatments

ABSTRACT The systematic understanding of how various antimicrobial agents are involved in controlling biofilms is essential in order to establish an effective strategy for biofilm control, since many antimicrobial agents are effective against planktonic cells but are ineffective when they are used against the same bacteria growing in a biofilm state. Three different antimicrobial agents (chlorine, silver, and tobramycin) and three different methods for the measurement of membrane integrity (plate counts, the measurement of respiratory activity with 5-cyano-2,3-ditolyl tetrazolium chloride [CTC] staining, and BacLight Live/Dead staining) were used along with confocal laser scanning microscopy (CLSM) and epifluorescence microscopy to examine the activities of the antimicrobials on biofilms in a comparative way. The three methods of determining the activities of the antimicrobials gave very different results for each antimicrobial agent. Among the three antimicrobials, tobramycin appeared to be the most effective in reducing the respiratory activity of biofilm cells, based upon CTC staining. In contrast, tobramycin-treated biofilm cells maintained their membrane integrity better than chlorine- or silver-treated ones, as evidenced by imaging by both CLSM and epifluorescence microscopy. Combined and sequential treatments with silver and tobramycin showed an enhanced antimicrobial efficiency of more than 200%, while the antimicrobial activity of either chlorine or tobramycin was antagonized when the agents were used in combination. This observation makes sense when the different oxidative reactivities of chlorine, silver, and tobramycin are considered.

3-D imaging of cells using a confocal laser scanning microscope and digital image processing

1988 ◽  
Vol 46 ◽  
pp. 96-97
Author(s):  
Thomas M. Jovin ◽  
Michel Robert-Nicoud ◽  
Donna J. Arndt-Jovin ◽  
Thorsten Schormann
Keyword(s):  
Laser Scanning ◽  
Confocal Laser Scanning Microscope ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Confocal Laser ◽  
Scanning Microscope ◽  
Laser Scanning Microscope ◽  
Biochemical Processes ◽  
Adjacent Structures

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.

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