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Biomedical subjects

D E Caldwell

Publications and source records attributed to D E Caldwell.

12 recordsLinked to original sources
PubMed1995

Lytic infection of Escherichia coli biofilms by bacteriophage T4.

Escherichia coli 3000 XIII formed biofilms on the surface of polyvinylchloride coupons in a modified Robbins device. Bacteriophage T4D+ infected cells in the biofilm and replicated. It is commonly held that bacteriophage cannot infect surface-attached bacteria (biofilms) because such bacteria are protected by an exopolymeric matrix that binds macromolecules and prevents their diffusion into the biofilm. To our knowledge this is the first observation that a bacteriophage can infect and multiply within cells growing as a biofilm.

Bacteriolysis↗
PubMed1992

Behavioral analysis of Vibrio parahaemolyticus variants in high- and low-viscosity microenvironments by use of digital image processing.

Digital image analysis and light microscopy were used to study and quantify the growth and behavior of two variants and selected flagellar mutants of Vibrio parahaemolyticus in glass flow cells under high- and low-viscosity conditions. The observations showed a series of surface-associated behaviors, including attachment, microcolony formation, migration, chemotactic movements, and aggregation, indicating a substantial degree of adaptive flexibility and multicellular behavior during growth of V. parahaemolyticus at interfaces.

Bacterial Adhesion↗
PubMed1991

Proteus mirabilis biofilm protection against struvite crystal dissolution and its implications in struvite urolithiasis.

Proteus mirabilis biofilm formation, struvite (MgNH4PO4.6H2O) crystal formation and dissolution in an artificial urine mixture were monitored using computer-enhanced microscopy (CEM) and a 1 x 3 mm. glass flow cell. Image analysis showed that P. mirabilis biofilm formation did not occur to any extent at macroenvironment flow rates greater than two mL/h (equivalent to a microenvironment flow rate of less than 5 microns./sec). Essentially, cells attached to glass surfaces, grew slowly and divided. Daughter cells were generally released directly into the medium where they could then presumably colonize other regions. Microcolonies formed by the adhesion of aggregates of cells from the medium, and over time grew into biofilms. Struvite crystallization due to urease activity and pH elevation above neutrality, was preceded by the deposition of organic matter on the glass surface, followed by the appearance of a number of tiny (one to two microns.) crystals. Crystals forming within a biofilm at low dilution rates took on a characteristic twinned or "X-shaped" appearance (crystal habit) indicative of a rapid growth rate. Those forming outside the biofilm took on a more tabular appearance reflecting their slower growth. When the macroenvironment flow rate of artificial urine (initial pH 5.8) in the glass flow cell was increased from two mL/h to four mL/h, struvite crystals not associated with biofilms dissolved within five to 10 min. Crystals entrapped within the P. mirabilis biofilm withstood flow rates up to 200 mL/h presumably due to the maintenance of an alkaline Mg-saturated microenvironment within the biofilm. These observations may suggest a mechanism by which struvite calculi can grow in spite of neutral or acidic urine pH and resist mild acidification therapy.

Bacterial Adhesion↗
PubMed1991

Optical sectioning of microbial biofilms.

Scanning confocal laser microscopy (SCLM) was used to visualize fully hydrated microbial biofilms. The improved rejection of out-of-focus haze and the increased resolution of SCLM made it preferable to conventional phase microscopy for the analysis of living biofilms. The extent of image improvement was dependent on the characteristics of individual biofilms and was most apparent when films were dispersed in three dimensions, when they were thick, and when they contained a high number of cells. SCLM optical sections were amenable to quantitative computer-enhanced microscopy analyses, with minimal interference originating from overlying or underlying cell material. By using SCLM in conjunction with viable negative fluorescence staining techniques, horizontal (xy) and sagittal (xz) sections of intact biofilms of Pseudomonas aeruginosa, Pseudomonas fluorescens, and Vibrio parahaemolyticus were obtained. These optical sections were then analyzed by image-processing techniques to assess the distribution of cellular and noncellular areas within the biofilm matrices. The Pseudomonas biofilms were most cell dense at their attachment surfaces and became increasingly diffuse near their outer regions, whereas the Vibrio biofilms exhibited the opposite trend. Biofilms consisting of different species exhibited distinctive arrangements of the major biofilm structural components (cellular and extracellular materials and space). In general, biofilms were found to be highly hydrated, open structures composed of 73 to 98% extracellular materials and space. The use of xz sectioning revealed more detail of biofilm structure, including the presence of large void spaces within the Vibrio biofilms. In addition, three-dimensional reconstructions of biofilms were constructed and were displayed as stereo pairs. Application of the concepts of architectural analysis to mixed- or pure-species biofilms will allow detailed examination of the relationships among biofilm structure, adaptation, and response to stress.

Image Processing, Computer-Assisted↗
PubMed1978

A Zoogloea sp. associated with blooms of Anabaena flos-aquae.

Bacteria were found attached to the heterocysts of Aphanizomenon flos-aquae and embedded within the mucilage of both anabaena flos-aquae and Microcystis aeruginosa in freshwater plankton. Electron microscopy of thin sections preceding the peak of an Anabaena flos-aquae bloom showed that the density of bacterial cells was 7.4 X 10(5) cells/ml in the planktonic macroenvironment and 2.6 X 10(11) cells/ml within the microenvironment of cyanobacterial mucilage. The bacteria occurred in aggregates and isolation required that these be dispersed by homogenizing at 50 000 rpm with glass beads. This procedure yielded a single bacterial isolate from blooms of Anabaena flos-aquae during 2 consecutive years. The isolate was flagellated, catalase- and oxidase-positive. Gram-negative, and rod-shaped to pleomorphic. Observation that the isolate required a pH greater than 8 for consistent growth, could not grow alone on liquid media but could grow alone on the corresponding solid media, could grow in liquid media only in the presence of Anabaena, formed tough mucilagenous colonies on solid media only in the presence of Anabaena extract, and rapidly assimilated but did not respire extracellular 14C-labelled organic matter produced by Anabaena suggested that the occurrence of the bacterium in cyanobacterial mucilage was not coincidental but reflected an obligatory bacterial requirement for the biological or physicochemical microenvironment of the mucilage. The bacterial isolate occurred in three growth forms. Either as a planktonic swarmer cell (which showed a positive chemotactic response to the cyanobacterium) embedded in cyanobacterial mucilage, or embedded in its own mucilage derived, in part, from a low molecular weight (below 1300) carbohydrate secreted by the cyanobacterium. These cultural, biochemical, and ecological characteristics suggest that the isolate is a new species in the genus Zoogloea and of potential importance in phytoplankton ecology.

Cyanobacteria↗
PubMed1977

Accessory pigment fluorescence for quantitation of photosynthetic microbial populations.

The in vivo fluorescence of the primary accessory pigments in purple bacteria (carotenoids), green bacteria (bacteriochlorophyll), green algae (chlorophyll), and cyanobacteria (phycocyanin) was found to be a linear function of cell concentration over three of four orders of magnitude. The lowest cell concentrations detectable were 10(4) cells/ml for procaryotes and 10(3) cells/ml for eucaryotes.

Bacteria↗
PubMed1976

Thermothrix thioparus gen. et sp. nov. a facultatively anaerobic facultative chemolithotroph living at neutral pH and high temperature.

Thermothrix thioparus gen. et ep. nov. occurs naturally in a New Mexico hot spring at a temperature of 74 degrees C, a pH of 7.0, and a HS- concentration of 1 mg/litre. The organism is gram-negative, non-motile, 0.5-1.0 X 3-20 mum, and forms cell chains up to 1 cm in length. The resulting filaments do not possess a sheath. Sulfur is deposited extracellularly. The organism was isolated using an autotrophic medium with HS- as the energy source and NO3- as the terminal electron acceptor. Anaerobically either NO2- or NO3- is required, NO2- is formed from NO3-, and no observable gas is evolved. Oxygen can also be used as the terminal electron acceptor, but growth is poor because of the decreased solubility of O2 at temperatures required for growth. Alternate energy sources used aerobically and anaerobically include hexose, HS-, SO3-, and S2O3=. The temperature optimum is 70-73 degrees C and growth occurs from 62 to 77 degrees C. The organism's thermal and physiological characteristics are compared to those of Bacillus stearothermophilus, Methanobacterium thermoautotrophicum, Sulfolobus acidocalderius, Thermus aquaticus, Thermus flavus, as well as Thiobacillus denitrificans, the latter being the only other facultatively anaerobic chemolithotroph which has been isolated and described.

Aerobiosis↗
PubMed1975

A morphological study of anaerobic bacteria from the hypolimnia of two Michigan lakes.

Dense populations of anaerobic bacteria were found sequentially layered below the thermocline in two eutrophic lakes in southwest Michigan. Phase and electron microscopy of whole cells and thin sections were used to reveal the in situ morphology of the dominant members of the community. The predominant chlorophyll-containing bacteria were identified on the basis of their morphology to be members of the genera Pelodictyon, Prosthecochloris, Clathrochloris, Chlorochromatium, Pelochromatium, Thiopedia, Thiocystis, Thiospirillum, and Chromatium. The natural morphology of these organisms is described and compared with the morphology of reported isolates, the morphology of unisolated genera was compared with previous descriptions of natural samples. Most of the organisms near the sediment-water interface and two from the upper hypolimnion have not been previously described. They have been divided into six distinct groups based on morphology; the morphological features of each group are presented. This approach, based on the morphological uniqueness of the procaryotes present, provides a satisfactory method for grouping members of the hypolimnetic community for ecological studies.

Anaerobiosis↗
PubMed1975

The structure of anaerobic bacterial communities in the hypolimnia of several Michigan lakes.

The structure of bacterial communities, the distribution of sulfide and oxygen, bacteriochlorophyll concentrations, and the temperature profile were determined for the anaerobic hypolimnia of two lakes in southern Michigan. Information from these studies, plus qualitative observations of two other lakes and two ponds over a 4-year period were used to correlate the spatial distribution of the populations, cell size, arrangement of photosynthetic vesicles or lamellae, presence of gas vacuoles or flagella, sulfur deposition, and environmental factors. On the basis of these results, three communities designated as A, B, and C were defined. The upper (A) community consisted of sequentially layered purple sulfur bacteria including two or more of the following genera: Thiopedia, Thiospirillum, Thiocystis, or Chromatium. The middle (B) community consisted of sequential layers of green bacteria from one or more of the following genera: Pelodictyon, Clathrochloris, Chlorochromatium, or Prosthecochloris. The lowest (C)community contained previously unreported gas-vacuolate colorless bacteria 0 to 0.7 m above the sediment. Microstratification (0.1- to 0.2-m layers) of populations was observed within the A and B communities.

Anaerobiosis↗