In-situ monitoring of Cryptosporidium parvum oocyst surface adhesion using ATR-FTIR spectroscopy.
ABSTRACT Surface chemistry and molecular interaction mechanisms of Cryptosporidium parvum oocysts with a ZnSe internal reflection element (IRE) surface were investigated as a function of pH and ionic strength in NaCl and CaCl(2) background electrolyte using in-situ ATR-FTIR spectroscopy. Since the surface properties of oocysts play an important role in adhesion behavior, the effects of surface modifications that are commonly employed to inactivate the pathogen for laboratory studies, including viable (control), formalin-, and heat-inactivation, were also examined. The ATR-FTIR spectra of oocyst surfaces exhibit amide, carboxylate, phosphate, and polysaccharide functional groups. Results indicate that changes in solution chemistry strongly impact oocyst adhesion behavior in aqueous systems. Increasing ionic strength from 1 to 100 mM or decreasing pH from 9.0 to 3.0 resulted in an increase in oocyst adhesion to the IRE surface as measured by IR absorbance. For equivalent ionic strength, the adhesion rate was found to be independent of CaCl(2) versus NaCl electrolyte solution, but was increased following formalin and heat treatments. This latter effect correlated with molecular changes reflected in spectral data. The ratio of amide I:amide II band intensities increased, and sugar ring vibrations at 1023 cm(-1) became sharper and more intense following formalin treatment. Similar changes in the polysaccharide region were observed following heat treatment, and protein secondary structure was also altered from mainly parallel beta-sheet to anti-parallel beta-sheet conformation.
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ABSTRACT: We investigated the effects of Shewanella putrefaciens cells and extracellular polymeric substances on the sorption of As(III) and As(V) to goethite, ferrihydrite, and hematite at pH 7.0. Adsorption of As(III) and As(V) at solution concentrations between 0.001 and 20 μM decreased by 10 to 45% in the presence of 0.3 g L(-1) EPS, with As(III) being affected more strongly than As(V). Also, inactivated Shewanella cells induced desorption of As(V) from the Fe(III)-(hydr)oxide mineral surfaces. ATR-FTIR studies of ternary As(V)-Shewanella-hematite systems indicated As(V) desorption concurrent with attachment of bacterial cells at the hematite surface, and showed evidence of inner-sphere coordination of bacterial phosphate and carboxylate groups at hematite surface sites. Competition between As(V) and bacterial phosphate and carboxylate groups for Fe(III)-(oxyhydr)oxide surface sites is proposed as an important factor leading to increased solubility of As(V). The results from this study have implications for the solubility of As(V) in the soil rhizosphere and in geochemical systems undergoing microbially mediated reduction and indicate that the presence of sorbed oxyanions may affect Fe-reduction and biofilm development at mineral surfaces.Environmental Science & Technology 03/2011; 45(7):2804-10. · 5.26 Impact Factor
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ABSTRACT: The protozoan parasites Giardia duodenalis, Cryptosporidium spp., and Toxoplasma gondii are pathogens that are resistant to a number of environmental factors and pose significant risks to public health worldwide. Their environmental transmission is closely governed by the physicochemical properties of their cysts (Giardia) and oocysts (Cryptosporidium and Toxoplasma), allowing their transport, retention, and survival for months in water, soil, vegetables, and mollusks, which are the main reservoirs for human infection. Importantly, the cyst/oocyst wall plays a key role in that regard by exhibiting a complex polymeric coverage that determines the charge and hydrophobic characteristics of parasites' surfaces. Interaction forces between parasites and other environmental particles may be, in a first approximation, evaluated following the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory of colloidal stability. However, due to the molecular topography and nano- to microstructure of the cyst/oocyst surface, non-DVLO hydrophobic forces together with additional steric attractive and/or repulsive forces may play a pivotal role in controlling the parasite behavior when the organism is subjected to various external conditions. Here, we review several parameters that enhance or hinder the adhesion of parasites to other particles and surfaces and address the role of fast-emerging techniques for mapping the cyst/oocyst surface, e.g., by measuring its topology and the generated interaction forces at the nano- to microscale. We discuss why characterizing these interactions could be a crucial step for managing the environmental matrices at risk of microbial pollution.Applied and Environmental Microbiology 12/2011; 78(4):905-12. · 3.95 Impact Factor
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ABSTRACT: The ability of microorganisms to adhere to abiotic surfaces and the potentialities of attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy have been exploited to study protonation and heavy metal binding events onto bacterial surfaces. This work represents the first attempt to apply on bacteria the recently developed method known as perfusion-induced ATR-FTIR difference spectroscopy. Such a technique allows measurement of even slight changes in the infrared spectrum of the sample, deposited as a thin layer on an ATR crystal, while an aqueous solution is perfused over its surface. Solutions at different pH have been used for inducing protonation/deprotonation of functional groups lying on the surface of Rhodobacter sphaeroides cells, chosen as a model system. The interaction of Ni(2+) with surface protonable groups of this microorganism has been investigated with a double-difference approach exploiting competition between nickel cations and protons. Protonation-induced difference spectra of simple model compounds have been acquired to guide band assignment in bacterial spectra, thus allowing identification of major components involved in proton uptake and metal binding. The data collected reveal that carboxylate moieties on the bacterial surface of R. sphaeroides play a role in extracellular biosorption of Ni(2+), establishing with this ion relatively weak coordinative bonds.Langmuir 03/2011; 27(7):3762-73. · 4.38 Impact Factor