Biomedical subjects

Alan W Decho

Publications and source records attributed to Alan W Decho.

7 recordsLinked to original sources

PubMed2005

The role of various dissolved organic matter forms on chlorpyrifos bioavailability to the estuarine bivalve Mercenaria mercenaria.

Dissolved organic matter (DOM) is comprised of a myriad of macromolecules with specific physical and chemical properties that may influence the bioavailability of hydrophobic pesticides to animals. This study was conducted to assess the role of various forms of DOM on the uptake and bioconcentration of the organophosphate insecticide chlorpyrifos (CHPY) to the bivalve Mercenaria mercenaria. Bivalves were exposed to DOM-free seawater (30 per thousand) or to seawater containing a single form of DOM. DOM forms included two filtrate fractions of natural salt-marsh sediment DOM (DOM-(<0.45 mum) and DOM-(<3 kDa)); natural purified humic (HA) and fulvic (FA) acids; and water soluble cyclic oligosaccharides alpha and beta cyclodextrins (CD-alpha and CD-beta). In (14)C-CHPY uptake and elimination experiments, juvenile bivalves were exposed to uniformly-labeled (14)C-CHPY and collected at time intervals during 48 h. The remaining bivalves were transferred to (14)C-CHPY-free elimination chambers with bivalve collection at time intervals over 144 h. Total uptake of (14)C-CHPY by bivalves in DOM-free seawater was >40% greater than in bivalves exposed to (14)C-CHPY in the presence of most DOM forms. These results are consistent with much faster (14)C-CHPY uptake rates estimated using a simple two parameter model. After the elimination period, bivalves exposed to DOM-free seawater had (14)C-CHPY body residue concentrations between 25% and 86% greater than bivalves in the presence of DOM forms. Experiments with larger bivalves showed that pulse-chase exposures with a 1.5 h exposure period to (14)C-CHPY was not long enough to detect differences in (14)C-CHPY tissue accumulation efficiencies across treatments. Our findings suggest that natural forms of DOM, at environmentally realistic organic carbon concentrations, reduced pesticide uptake and bioconcentration, consistent with much lower uptake rates relative to bivalves exposed to (14)C-CHPY in the absence of DOM. Interestingly, at the tested organic carbon concentrations CD-alpha and CD-beta did not reduce (14)C-CHPY bioconcentration in M. mercenaria.

Analysis of Variance↗

PubMed2004

Using geographical information techniques to quantify the spatial structure of endolithic boring processes within sediment grains of marine stromatolites.

Marine stromatolites are generated through the interactions of environmental parameters and specific microbial processes. The activities of endolithic bacteria, that bore canals through calcium carbonate (CaCO(3)) sand grains (ooids) and reprecipitate the CaCO(3) as a single layer (i.e. micritic laminae) are especially important in the longer term stability of the stromatolite macrostructure. Image analysis and classification approaches have been used previously, but only seldom as a quantitative microscopic tool. Here, we develop a new approach that enables the quantification of microscale (i.e. micrometers to millimeters) spatial structure within marine stromatolites. To demonstrate our approach, images were acquired from two different layers of a stromatolite: "orange layers", where microboring of canals within ooids was relatively abundant, and "white layers" where microboring was greatly reduced or lacking. Images were then transformed into spatial maps. Computation of canal and ooid grain areas within each image was conducted and statistically compared between replicate samples from the two stromatolite layers. This allowed quantification of the areas of ooid grains that were microbored. Based on our results, we suggest that our method could be widely applicable to sedimentary environments, and other areas of fundamental research.

Bacteria↗

PubMed2003

Development of an indirect competitive enzyme-linked immunosorbent assay to detect extracellular polymeric substances (EPS) secreted by the marine stromatolite-forming cyanobacteria, Schizothrix sp.

An indirect competitive enzyme-linked immunosorbent assay was developed using polyclonal antibody to detect extracellular polymeric secretions (EPS) produced by the marine stromatolite-forming cyanobacteria, Schizothrix sp. The cross-reactivity of this assay with other EPSs and polymers were low (< 0.5%). This assay can detect Schizothrix sp. EPS as low as 0.5 ng/mL. Intra-assay and inter-assay comparisons showed that coefficient variations were low, ranging from 3.34 to 10.30% and from 6.30 to 12.8%, respectively, for standards between 2 and 1000 ng/mL. Also, the seawater matrix effect was negligible. Our results indicated that this assay is a useful tool for quantification of Schizothrix sp. EPS in a range of ng/mL.

Cross Reactions↗

PubMed2003

Bioavailability of the organophosphorous insecticide chlorpyrifos to the suspension-feeding bivalve, Mercenaria mercenaria, following exposure to dissolved and particulate matter.

Under estuarine conditions, hydrophobic pesticides undergo physical/chemical partitioning into dissolved and particulate fractions of sediments. Our experimental study using 14C-chlorpyrifos (14C-CHPY) showed that sorptive associations of this common insecticide with particulate (>0.2 microm) and dissolved fractions (<0.2 microm) of sediments strongly influenced uptake, tissue absorption, and elimination in the estuarine bivalve Mercenaria mercenaria. Bivalves were fed algae mixed with particulate and dissolved fractions of 14C-CHPY-contaminated estuarine sediment particles, components of estuarine sediments (silica, humic acids), and a naturally occurring food source (phytoplankton) to quantify relative uptake, tissue absorption efficiencies, and elimination of 14C-CHPY. Measurements of 14C-CHPY tissue absorption efficiencies (AE%) indicated that particle source qualities influenced much of the insecticide bioavailability. Mean tissue AE% of 14C-CHPY associated with different particulate fractions ranged from 23 to 31%, while uptake from the dissolved/colloidal forms resulted in low (7-17%) AE%. Our data indicated that 14C-CHPY is more likely to bioaccumulate through ingestion of contaminated particulate material than through filtration/ingestion of dissolved/colloidal material. The high selectivity and digestibility of algae particles by M. mercenaria, a potentially ingested food source, may play an important role in the bioavailability of chlorpyrifos in this animal.

Animals↗

PubMed2002

In situ microspatial imaging using two-photon and confocal laser scanning microscopy of bacteria and extracellular polymeric secretions (EPS) within marine stromatolites.

The combination of a hydrophilic embedding resin, Nanoplast, with fluorescent probes, and subsequent imaging using two-photon and confocal laser scanning microscopy (2P-LSM and CLSM) has allowed in imaging of the in situ microspatial arrangements of microbial cells and their extracellular polymeric secretion (EPS) within marine stromatolites. Optical sectioning by 2P-LSM and CLSM allowed imaging of endolithic cyanobacteria cells, Solentia sp., seen within carbonate sand grains. 2P-LSM allowed very clear imaging with a high resolution of bacteria using DAPI, which normally require UV excitation and reduced photo-bleaching of fluorescent probes.

Journal Article↗

PubMed2002

Isolation and biochemical characterization of extracellular polymeric secretions (EPS) from modern soft marine stromatolites (Bahamas) and its inhibitory effect on CaCO3 precipitation.

Bahamian soft marine stromatolites consist of cyanobacterial biofilms and carbonate sand grains (ooids) embedded in their extracellular polymeric secretions (EPS). EPS were isolated from natural marine stromatolites and the laboratory cultured stromatolite forming cyanobacterium isolate Schizothix sp. Laboratory investigations were conducted to examine biochemical characteristics and the role of EPS in the inhibition of CaCO3 precipitation. EPS consisted of acid polysaccharides and proteins. SDS-PAGE and amino acid analysis suggested that EPS from both soft marine stromatolite and Schizothrix sp. mat contained small proteins (38 kD and 45 kD) enriched in aspartic acid and glutamic acid. Also, immuno blotting suggested that natural EPS contain high molecular weight acid polysaccharide (500 k) which may represent cross-linked products of laboratory cultured Schizothrix sp. acid polysaccharide (300 k). EPS from both soft marine stromatolite and laboratory cultured Schizothrix sp. inhibited CaCO3 precipitation in vitro, as determined using pH drift assays examining pH decrease which occur in response to CaCO3 precipitation. PH drift assays of enzymatically and chemically modified EPS isolated from soft marine stromatolite and laboratory cultured Schizothrix sp. indicated that both uronic acids and protein fractions may be involved in the inhibition of CaCO3 precipitation.

Amino Acids↗

PubMedDate not supplied

Reconstruction and computation of microscale biovolumes using geographical information systems: potential difficulties.

Biofilms are bacterial colonies enveloped in a matrix of extracellular polymeric secretions. Confocal scanning laser microscopy has been used in conjunction with different image analysis techniques to investigate the structure of biofilms. A major goal is to reconstitute the three-dimensional structure of biofilms, and compute or estimate the biovolumes. Our previous research focused on the utilization of remote sensing techniques and Geographical Information Systems for quantitative analyses of confocal images. The present study investigates potential problems in microbial imaging, and uses two approaches, the program COMSTAT and a Geographical Information Systems-based method, to reconstitute three-dimensional structures and estimate biovolumes. Volumes of thirty fluorescent polymeric microspheres with a known diameter were estimated and used as a ground truth, to statistically compare both methods. In a next step, the two approaches were used to estimate the biovolume of a section through a Pseudomonas aeruginosa biofilm. Difficulties were encountered in image acquisition due to the optical properties of the microbeads. Our results indicate that the Geographical Information Systems approach produced results consistent with the existing COMSTAT approach, and close to theoretical values, despite many problems inherent to each phase of this process. Also, the image classification process encountered several limitations. It is suggested that the unique constraints of the microscopic world may generate additional problems, especially related to image classification.

Biofilms↗