Environmental implications of one-century COPRs evolution in a single industrial site: From leaching impact to sustainable remediation of CrVI polluted groundwater.

The Stoppani factory manufactured chromium for more than one century, dumping millions of tons of Chromite Ore Processing Residues (COPRs) over decades. The massive presence of COPRs resulted in an intense CrVI leaching and consequent contamination of percolating groundwater. The site offers a unique opportunity to follow COPRs evolution from the primary roasting process to the aged Cr-bearing mineral phases. Herein, new insights on COPRs mineralogy evolution and their role in CrVI release are provided by a dry sample preparation protocol, coupled with in-depth multi-technique characterization. Besides typical COPRs mineral assemblages, highly soluble Na2CrO4 and the first evidence of crocoite (PbCrO4) in a COPR contaminated site are revealed. Selective extraction experiments confirmed a strong reactivity for Cr-bearing minerals as confirmed by concentrations as high as 375 mg L-1 of leached CrVI. The mineralogical approach was combined with a nanotechnological solution for CrVI wastewater remediation. The application of naked colloidal maghemite (γ-Fe2O3) nanoparticles (SAMNs) on the complex industrial wastewater, led to > 90% CrVI removal, either under acidic or in-situ conditions. The present case study of a highly polluted site, ranging from mineral characterization to wastewater remediation, highlights the use of multidisciplinary approaches to cope with complex environmental issues.

Role of carboxylic group pattern on protein surface in the recognition of iron oxide nanoparticles: A key for protein corona formation.

The knowledge of protein-nanoparticle interplay is of crucial importance to predict the fate of nanomaterials in biological environments. Indeed, protein corona on nanomaterials is responsible for the physiological response of the organism, influencing cell processes, from transport to accumulation and toxicity. Herein, a comparison using four different proteins reveals the existence of patterned regions of carboxylic groups acting as recognition sites for naked iron oxide nanoparticles. Readily interacting proteins display a distinctive surface distribution of carboxylic groups, recalling the geometric shape of an ellipse. This is morphologically complementary to nanoparticles curvature and compatible with the topography of exposed FeIII sites laying on the nanomaterial surface. The recognition site, absent in non-interacting proteins, promotes the nanoparticle harboring and allows the formation of functional protein coronas. The present work envisages the possibility of predicting the composition and the biological properties of protein corona on metal oxide nanoparticles.

Smart synthetic maghemite nanoparticles with unique surface properties encode binding specificity toward AsIII.

Pristine ɣ-Fe2O3 nanoparticles, called surface active maghemite nanoparticles (SAMNs) display unprecedented colloidal stability and specific binding properties. Herein, the interactions of SAMNs with AsV and AsIII as surface molecular probes were comparatively studied. Thermodynamic and kinetic characterizations, along with chemical and structural analysis of SAMN@As complexes, evidenced two distinct binding modalities. Arsenite, emerged as an elective and specific ligand for SAMNs, whereas arsenate adsorption was more labile, pH dependent and ruled by different binding possibilities. In particular, AsIII oxyacid exclusively interacts through inner-sphere coordination occupying available surface crystal positions resembling a key-lock fitting, while AsV leads to both outer-sphere and inner-sphere complexes. Noteworthy, discrimination between AsV and AsIII was never reported for nanostructured maghemite evidencing the importance of synthetic route on surface properties of the nanomaterial. The present report, besides enriching the chemistry of nanosized iron oxides, suggests SAMNs application for the remediation of water contaminated by AsIII, the most threatening As species in water.

Enzyme Immobilization on Maghemite Nanoparticles with Improved Catalytic Activity: An Electrochemical Study for Xanthine.

Generally, enzyme immobilization on nanoparticles leads to nano-conjugates presenting partially preserved, or even absent, biological properties. Notwithstanding, recent research demonstrated that the coupling to nanomaterials can improve the activity of immobilized enzymes. Herein, xanthine oxidase (XO) was immobilized by self-assembly on peculiar naked iron oxide nanoparticles (surface active maghemite nanoparticles, SAMNs). The catalytic activity of the nanostructured conjugate (SAMN@XO) was assessed by optical spectroscopy and compared to the parent enzyme. SAMN@XO revealed improved catalytic features with respect to the parent enzyme and was applied for the electrochemical studies of xanthine. The present example supports the nascent knowledge concerning protein conjugation to nanoparticle as a means for the modulation of biological activity.

Electrocatalytic nanostructured ferric tannate as platform for enzyme conjugation: Electrochemical determination of phenolic compounds.

A shell of nanostructured ferric tannates was spontaneously developed on the surface of naked maghemite nanoparticles (SAMNs, the core) by a simple wet reaction with tannic acid (TA). The as obtained core-shell nanomaterial (SAMN@TA) displays specific electrocatalytic and surface properties, which significantly differ from parent maghemite. Thanks to the known proclivity of TA to interact with proteins, SAMN@TA was proposed as a support for the direct immobilization of an enzyme. A ternary functional nanobioconjugate (SAMN@TA@TvL) was successfully self-assembled by incubating laccase from Trametes versicolor (TvL) and SAMN@TA. The SAMN@TA@TvL hybrid was kinetically characterized with respect to the native enzyme and applied for building an easy-to-use analytical device for the detection of polyphenols. The electrochemical biosensor allowed the determination of polyphenols by square wave voltammetry in mixed water-methanol solutions. The system sensitivity was 868.9 ±â€¯1.9nA µM-1, the LOD was 81 nM and the linearity range was comprised between 100 nM and 10 µM. The proposed approach was successfully applied to detect phenolics in blueberry extracts as real samples. Results suggest that SAMN@TA could be a promising, low cost and versatile tool for the creation of nano-bio-conjugates aimed at the development of new electrochemical sensing platforms.

Biologically safe colloidal suspensions of naked iron oxide nanoparticles for in situ antibiotic suppression.

A category of naked maghemite nanoparticles (γ-Fe2O3), named surface active maghemite nanoparticles (SAMNs), is characterized by biological safety, high water colloidal stability and a surface chemistry permitting the binding of ligands. In the present study, the interaction between SAMNs and an antibiotic displaying chelating properties (oxytetracycline, OxyTC) was extensively structurally and magnetically characterized. OxyTC emerged as an ideal probe for providing insights into the colloidal properties of SAMNs. At the same time, SAMNs turned out as an elective tool for water remediation from OxyTC. Therefore, a dilute colloidal suspension of SAMNs was used for the removal of OxyTC in large volume tanks where, to simulate a real in situ application, a population of zebrafish (Danio rerio) was introduced. Interestingly, SAMNs led to the complete removal of the drug without any sign of toxicity for the animal model. Moreover, OxyTC immobilized on SAMNs surface resulted safe for sensitive Escherichia coli bacteria strain. Thus, SAMNs were able to recover the drug and to suppress its antibiotic activity envisaging their feasibility as competitive option for water remediation from OxyTC in more nature related scenarios. The present contribution stimulates the use of novel smart colloidal materials to cope with complex environmental issues.

Self-assembly of chlorin-e6 on γ-Fe2O3 nanoparticles: Application for larvicidal activity against Aedes aegypti.

Aedes aegypti mosquitos are widespread vectors of several diseases and their control is of primary importance for biological and environmental reasons, and novel safe insecticides are highly desirable. An eco-friendly photosensitizing magnetic nanocarrier with larvicidal effects on Aedes aegypti was proposed. The innovative core-shell hybrid nanomaterial was synthesized by combining peculiar magnetic nanoparticles (called Surface Active Maghemite Nanoparticles - SAMNs, the core) and chlorin-e6 as photosensitizer (constituting the shell) via self-assembly in water. The hybrid nanomaterial (SAMN@chlorin) was extensively characterized and tested for the photocidal activity on larvae of Aedes aegypti. The SAMN@chlorin core-shell nanohybrid did not present any toxic effect in the dark, but, upon light exposure, showed a higher photocidal activity than free chlorin-e6. Moreover, the eco-toxicity of SAMN@chlorin was determined in adults and neonates of Daphnia magna, where delayed toxicity was observed only after prolonged (≥4 h) exposure to intense light, on the green alga Pseudokirchneriella subcapitata and on the duckweed Lemna minor on which no adverse effects were observed. The high colloidal stability, the physico-chemical robustness and the magnetic drivability of the core-shell SAMN@chlorin nanohybrid, accompanied by the high photocidal activity on Aedes aegypti larvae and reduced environmental concerns, can be proposed as a safe alternative to conventional insecticides.

Enzyme self-assembly on naked iron oxide nanoparticles for aminoaldehyde biosensing.

The preservation of enzymatic activity is a fundamental requirement for exploiting hybrid nano-bio-conjugates, and the control over protein-nanoparticle interactions, leading to stable and catalytically active hybrids, represents the key for designing new biosensing platforms. In this scenario, surface active maghemite nanoparticles (SAMNs) represent a new class of naked magnetic nanoparticles, displaying peculiar electrocatalytic features and the ability to selectively bind proteins. Recombinant aminoaldehyde dehydrogenase from tomato (SlAMADH1) was used as a model protein, and successfully immobilized by self-assembly on the surface of naked SAMNs, where its enzymatic activity resulted preserved for more than 6 months. The hybrid nanomaterial (SAMN@SlAMADH1) was characterized by UV-Vis spectroscopy, mass spectrometry, and TEM microscopy, and applied for the development of a biosensor for the determination of aminoaldehydes in alcoholic beverages. Measurements were carried out in a low volume electrochemical flow cell comprising a SAMN modified carbon paste electrode for the coulometric determination of the NADH produced during the enzymatic catalysis. The present findings, besides representing the first example of an electrochemical biosensor for aminoaldehydes in an alcoholic matrix, open the door to the use of immobilized enzymes on naked metal oxides nanomaterials for biosensing.

Stealth Iron Oxide Nanoparticles for Organotropic Drug Targeting.

The ability of peculiar iron oxide nanoparticles (IONPs) to evade the immune system was investigated in vivo. The nanomaterial was provided directly into the farming water of zebrafish ( Danio rerio) and the distribution of IONPs and the delivery of oxytetracycline (OTC) was studied evidencing the successful overcoming of the intestinal barrier and the specific and prolonged (28 days) organotropic delivery of OTC to the fish ovary. Noteworthy, no sign of adverse effects was observed. In fish blood, IONPs were able to specifically bind apolipoprotein A1 (Apo A1) and molecular modeling showed the structural analogy between the IONP@Apo A1 nanoconjugate and high-density lipoprotein (HDL). Thus, the preservation of the biological identity of the protein suggests a plausible explanation of the observed overcoming of the intestinal barrier, of the great biocompatibity of the nanomaterial, and of the prolonged drug delivery (benefiting of the lipoprotein transport route). The present study promises novel and unexpected stealth materials in nanomedicine.

Versatile nano-platform for tailored immuno-magnetic carriers.

Custom immuno-magnetic devices are desirable tools for biomedical and biotechnological applications. Herein, surface active maghemite nanoparticles (SAMNs) are proposed as a versatile platform for developing tailored immuno-magnetic nano-carriers by simple wet reactions. Two examples for conjugating native and biotinylated antibodies were presented along with their successful applications in the recognition of specific foodborne pathogens. Nanoparticles were functionalized with rhodamine B isothiocyanate (RITC), leading to a fluorescent nano-conjugate, and used for binding anti-Campylobacter fetus antibodies (SAMN@RITC@Anti-Cf). The microorganism was selectively captured in the presence of two other Campylobacter species (C. jejuni and C. coli), as verified by PCR. Alternatively, SAMNs were modified with avidin, forming a biotin-specific magnetic nano-carrier and used for the immobilization of biotinylated anti-Listeria monocytogenes antibodies (SAMN@avidin@Anti-Lm). This immuno-magnetic carrier was integrated in piezoelectric quartz crystal microbalance (QCM) sensor for the detection of L. monocytogenes in milk, showing a detection limit of 3 bacterial cells. The present work presents a new category of customized immuno-magnetic nano-carriers as a competitive option for suiting specific applications. Graphical abstract ᅟ.

The surface reactivity of iron oxide nanoparticles as a potential hazard for aquatic environments: A study on Daphnia magna adults and embryos.

Nano-ecotoxicology is extensively debated and nanomaterial surface reactivity is an emerging topic. Iron oxide nanoparticles are widely applied, with organic or inorganic coatings for stabilizing their suspensions. Surface active maghemite nanoparticles (SAMNs) are the unique example of naked iron oxide displaying high colloidal and structural stability in water and chemical reactivity. The colloidal behavior of SAMNs was studied as a function of the medium salinity and protocols of acute and chronic toxicity on Daphnia magna were consequently adapted. SAMN distribution into the crustacean, intake/depletion rates and swimming performances were evaluated. No sign of toxicity was detected in two model organisms from the first trophic level (P. subcapitata and L. minor). In D. magna, acute EC50 values of SAMN was assessed, while no sub-lethal effects were observed and the accumulation of SAMNs in the gut appeared as the sole cause of mortality. Fast depuration and absence of delayed effects indicated no retention of SAMNs within the organism. In spite of negligible toxicity on D. magna adults, SAMN surface reactivity was responsible of membrane bursting and lethality on embryos. The present study offers a contribution to the nascent knowledge concerning the impact of nanoparticle surface reactivity on biological interfaces.

Endogenous and food-derived polyamines: determination by electrochemical sensing.

Polyamines (PAs) are involved in a variety of fundamental physio-pathologic processes. The concentration of these polycations in organs and tissues depends on their endogenous production and oxidation rates, and on their intake from foods. Besides being largely accepted as markers for the progress of several pathologies, PAs may exert themselves different effects on humans, ranging from being positive to be drastically detrimental depending on the organism conditions. Thus, if the determination of polyamines content in tissue samples is of great importance as they could be indicators of several diseases, their quantification in food is fundamental for modulating the diet to respond to a specific human health status. Thus, the determination of PA content in food is increasingly urgent. Standard analytical methods for polyamine quantification are mainly based on chromatography, where high-performance liquid chromatography and gas chromatography are the most often used, involving pre-column or post-column derivatization techniques. Driven by the growing need for rapid in situ analyses, electrochemical biosensors, comprising various combinations of different enzymes or nanomaterials for the selective bio-recognition and detection, are emerging as competitors of standard detection systems. The present review is aimed at providing an up-to-date overview on the recent progresses in the development of sensors and biosensors for the detection of polyamines in human tissues and food samples. Basic principles of different electrochemical (bio)sensor formats are reported and the applications in human tissues and in foods was evidenced.

Antimicrobial and magnetically removable tannic acid nanocarrier: A processing aid for Listeria monocytogenes treatment for food industry applications.

An innovative core-shell nanocarrier, combining the magnetism of surface active maghemite nanoparticles (SAMNs, the core) and tannic acid (TA, the shell) was self-assembled by simple incubation in water. Due to the drastic reorganization of SAMN surface, the prepared magnetic nanocarrier (SAMN@TA) resulted as one of the most robust nanomaterial bearing TA to date. Nevertheless, the ferric tannates network, constituting the SAMN@TA shell, and the free tannic acid display comparable chemical behavior. The antimicrobial properties of SAMN@TA were tested on Listeria monocytogenes in comparison with free TA, showing similar bacteriostatic effects at relatively low concentrations. Besides the preservation of the TA inhibitory activity toward L. monocytogenes, the possibility of being magnetically removed leaving no residues into the matrix makes this nanocarrier an innovative processing aid for surface treatments. Thus, SAMN@TA can be used as an effective, low-cost and environmentally friendly antimicrobial nanomaterial for the food industry applications.

Analysis of hard protein corona composition on selective iron oxide nanoparticles by MALDI-TOF mass spectrometry: identification and amplification of a hidden mastitis biomarker in milk proteome.

Surface active maghemite nanoparticles (SAMNs) are able to recognize and bind selected proteins in complex biological systems, forming a hard protein corona. Upon a 5-min incubation in bovine whey from mastitis-affected cows, a significant enrichment of a single peptide characterized by a molecular weight at 4338 Da originated from the proteolysis of aS1-casein was observed. Notably, among the large number of macromolecules in bovine milk, the detection of this specific peptide can hardly be accomplished by conventional analytical techniques. The selective formation of a stable binding between the peptide and SAMNs is due to the stability gained by adsorption-induced surface restructuration of the nanomaterial. We attributed the surface recognition properties of SAMNs to the chelation of iron(III) sites on their surface by sterically compatible carboxylic groups of the peptide. The specific peptide recognition by SAMNs allows its easy determination by MALDI-TOF mass spectrometry, and a threshold value of its normalized peak intensity was identified by a logistic regression approach and suggested for the rapid diagnosis of the pathology. Thus, the present report proposes the analysis of hard protein corona on nanomaterials as a perspective for developing fast analytical procedures for the diagnosis of mastitis in cows. Moreover, the huge simplification of proteome complexity by exploiting the selectivity derived by the peculiar SAMN surface topography, due to the iron(III) distribution pattern, could be of general interest, leading to competitive applications in food science and in biomedicine, allowing the rapid determination of hidden biomarkers by a cutting edge diagnostic strategy. Graphical abstract The topography of iron(III) sites on surface active maghemite nanoparticles (SAMNs) allows the recognition of sterically compatible carboxylic groups on proteins and peptides in complex biological matrixes. The analysis of hard protein corona on SAMNs led to the determination of a biomarker for cow mastitis in milk by MALDI-TOF mass spectrometry.

New Perspectives on Biomedical Applications of Iron Oxide Nanoparticles.

Iron oxide nanomaterials are considered promising tools for improved therapeutic efficacy and diagnostic applications in biomedicine. Accordingly, engineered iron oxide nanomaterials are increasingly proposed in biomedicine, and the interdisciplinary researches involving physics, chemistry, biology (nanotechnology) and medicine have led to exciting developments in the last decades. The progresses of the development of magnetic nanoparticles with tailored physico-chemical and surface properties produced a variety of clinically relevant applications, spanning from magnetic resonance imaging (MRI), drug delivery, magnetic hyperthermia, to in vitro diagnostics. Notwithstanding the wellknown conventional synthetic procedures and their wide use, along with recent advances in the synthetic methods open the door to new generations of naked iron oxide nanoparticles possessing peculiar surface chemistries, suitable for other competitive biomedical applications. New abilities to rationally manipulate iron oxides and their physical, chemical, and biological properties, allow the emersion of additional possibilities for designing novel nanomaterials for theranostic applications.

Covalently bound DNA on naked iron oxide nanoparticles: Intelligent colloidal nano-vector for cell transfection.

BACKGROUND: Conversely to common coated iron oxide nanoparticles, novel naked surface active maghemite nanoparticles (SAMNs) can covalently bind DNA. Plasmid (pDNA) harboring the coding gene for GFP was directly chemisorbed onto SAMNs, leading to a novel DNA nanovector (SAMN@pDNA). The spontaneous internalization of SAMN@pDNA into cells was compared with an extensively studied fluorescent SAMN derivative (SAMN@RITC). Moreover, the transfection efficiency of SAMN@pDNA was evaluated and explained by computational model. METHODS: SAMN@pDNA was prepared and characterized by spectroscopic and computational methods, and molecular dynamic simulation. The size and hydrodynamic properties of SAMN@pDNA and SAMN@RITC were studied by electron transmission microscopy, light scattering and zeta-potential. The two nanomaterials were tested by confocal scanning microscopy on equine peripheral blood-derived mesenchymal stem cells (ePB-MSCs) and GFP expression by SAMN@pDNA was determined. RESULTS: Nanomaterials characterized by similar hydrodynamic properties were successfully internalized and stored into mesenchymal stem cells. Transfection by SAMN@pDNA occurred and GFP expression was higher than lipofectamine procedure, even in the absence of an external magnetic field. A computational model clarified that transfection efficiency can be ascribed to DNA availability inside cells. CONCLUSIONS: Direct covalent binding of DNA on naked magnetic nanoparticles led to an extremely robust gene delivery tool. Hydrodynamic and chemical-physical properties of SAMN@pDNA were responsible of the successful uptake by cells and of the efficiency of GFP gene transfection. GENERAL SIGNIFICANCE: SAMNs are characterized by colloidal stability, excellent cell uptake, persistence in the host cells, low toxicity and are proposed as novel intelligent DNA nanovectors for efficient cell transfection.

Colloidal Surface Active Maghemite Nanoparticles for Biologically Safe Cr(VI) Remediation: from Core-Shell Nanostructures to Pilot Plant Development.

The present study is aimed at the exploration of achievable improvements for Cr(VI) ex situ and in situ water remediation by using novel naked colloidal maghemite (γ-Fe2 O3 ) nanoparticles (surface active maghemite nanoparticles, SAMNs). The reliability of SAMNs for Cr(VI) binding and removal was demonstrated, and SAMN@Cr(VI) complex was characterized, as well as the covalent nature of the absorption was unequivocally proved. SAMNs were structurally and magnetically well conserved after Cr(VI) binding. Thus, in consideration of their affinity for Cr(VI) , SAMNs were exploited in a biological model system, mimicking a real in situ application. The assay evidenced a progressive reduction of revertant colonies of Salmonella typhimurium TA100 strain, as maghemite nanoparticles concentration increased, till the complete suppression of Cr(VI) mutagen effect. Finally, an automatic modular pilot system for continuous magnetic removal and recovery of Cr(VI) from water is proposed. SAMNs, thanks to their colloidal, binding, and catalytic properties, represent a promising tool as a reliable nanomaterial for water remediation by Cr(VI) .

Electrocatalytic Nanostructured Ferric Tannates: Characterization and Application of a Polyphenol Nanosensor.

A novel core-shell hybrid nanomaterial composed of peculiar maghemite nanoparticles (surface-active maghemite nanoparticles (SAMNs)) as the core and tannic acid (TA) as the shell was developed by self-assembly of ferric tannates onto the surface of SAMNs by simple incubation in water. The hybrid nanomaterial (SAMN@TA) was characterized by using UV/Vis, FTIR, and Mössbauer spectroscopies, magnetization measurements, and X-ray powder diffraction, which provide evidence of a drastic reorganization of the iron oxide surface upon reaction with TA and the formation of an outer shell that consists of a cross-linked network of ferric tannates. According to a Langmuir isotherm analysis, SAMN@TA offers one of most stable iron complexes of TA reported in the literature to date. Moreover, SAMN@TA was characterized by using electrical impedance spectroscopy, voltammetry, and chronoamperometry. The nanostructured ferric tannate interface showed improved conductivity and selective electrocatalytic activity toward the oxidation of polyphenols. Finally, a carbon-paste electrode modified with SAMN@TA was used for the determination of polyphenols in blueberry extracts by square-wave voltammetry.

Alkaline Water and Longevity: A Murine Study.

The biological effect of alkaline water consumption is object of controversy. The present paper presents a 3-year survival study on a population of 150 mice, and the data were analyzed with accelerated failure time (AFT) model. Starting from the second year of life, nonparametric survival plots suggest that mice watered with alkaline water showed a better survival than control mice. Interestingly, statistical analysis revealed that alkaline water provides higher longevity in terms of "deceleration aging factor" as it increases the survival functions when compared with control group; namely, animals belonging to the population treated with alkaline water resulted in a longer lifespan. Histological examination of mice kidneys, intestine, heart, liver, and brain revealed that no significant differences emerged among the three groups indicating that no specific pathology resulted correlated with the consumption of alkaline water. These results provide an informative and quantitative summary of survival data as a function of watering with alkaline water of long-lived mouse models.

Enlightening mineral iron sensing in Pseudomonas fluorescens by surface active maghemite nanoparticles: Involvement of the OprF porin.

BACKGROUND: Mineral iron(III) recognition by bacteria is considered a matter of debate. The peculiar surface chemistry of novel naked magnetic nanoparticles, called SAMNs (surface active maghemite nanoparticles) characterized by solvent exposed Fe(3+) sites on their surface, was exploited for studying mineral iron sensing in Pseudomonas fluorescens. METHODS: SAMNs were applied for mimicking Fe(3+) ions in solution, acting as magnetically drivable probes to evaluate putative Fe(3+) recognition sites on the microorganism surface. Culture broths and nano-bio-conjugates were characterized by UV-Vis spectroscopy and mass spectrometry. RESULTS: The whole heritage of a membrane porin (OprF) of P. fluorescens Ps_22 cells was recognized and firmly bound by SAMNs. The binding of nanoparticles to OprF porin was correlated to a drastic inhibition of a siderophore (pyoverdine) biosynthesis and to the stimulation of the production and rate of formation of a secondary siderophore. The analysis of metabolic pathways, based on P. fluorescens Ps_22 genomic information, evidenced that this putative secondary siderophore does not belong to a selection of the most common siderophores. CONCLUSIONS: In the scenario of an adhesion mechanism, it is plausible to consider OprF as the biological component deputed to the mineral iron sensing in P. fluorescens Ps_22, as well as one key of siderophore regulation. GENERAL SIGNIFICANCE: The present work sheds light on mineral iron sensing in microorganisms. Peculiar colloidal naked iron oxide nanoparticles offer a useful approach for probing the adhesion of bacterial surface on mineral iron for the identification of the specific recognition site for this iron uptake regulation in microorganisms.