Silver Resistance Research Articles

Assessment of antimicrobial effects of food contact materials containing silver on growth of Salmonella Typhimurium

Food contact materials containing antibacterial properties are progressively appearing in the market. Items intended to provide antimicrobial effects such as increased shelf life and food safety are incorporating silver materials during the manufacture of such products. This study examined the total silver content, release capacity, and antibacterial activity of three different silver-containing food contact materials: plastic food storage containers, a plastic cutting board, and food wrapping paper. Silver content and release were determined by Inductively Coupled Plasma Mass Spectrometry, and the results showed that, although the amount of silver in each product was similar, migration varied considerably with kind of material and simulant choice. Antimicrobial effect was tested by measuring the growth of Salmonella Typhimurium during or after exposure to the different food contact materials. The results showed that the food storage containers and wrapping paper delayed the growth of S. Typhimurium under certain conditions, but that these effects were short-lived. The strain of S. Typhimurium used in this study was found to be negative for the presence of tested silver resistance genes. The results of this study suggest that a thorough investigation should be required to show/claim the efficacy of silver-containing food contact materials for food safety purposes.

The relation of structure and dispersion to amorphous silicon silver thin films

Silver/amorphous silicon (Ag/a-Si) memristor has attracted extensive attention as a candidate for next-generation nonvolatile memories. The optical constant variations of a-Si layer during switching procedure, which are the key for Ag/a-Si memristor to be used in optical applications, have not been clarified yet. To take a further understanding on the optical feathers of Ag/a-Si memristor, here we report the microstructure, resistivity and dispersion relation of amorphous silicon silver (a-Si1−xAgx) thin films prepared by co-sputtering. Ag nanocrystals are observed in the films with size similar to the Ag filament in memristor, and the obtained resistivities of the films march the typical ON/OFF ratio of memristor. Furthermore, both the film's refraction index and extinction coefficient increase with Ag concentration, suggesting that Ag/a-Si memristor combining with silicon waveguide could be applied in optical switch at 1300 and 1550nm.

Electrical four-point probing of spherical metallic thin films coated onto micron sized polymer particles

Micron-sized metal-coated polymer spheres are frequently used as filler particles in conductive composites for electronic interconnects. However, the intrinsic electrical resistivity of the spherical thin films has not been attainable due to deficiency in methods that eliminate the effect of contact resistance. In this work, a four-point probing method using vacuum compatible piezo-actuated micro robots was developed to directly investigate the electric properties of individual silver-coated spheres under real-time observation in a scanning electron microscope. Poly(methyl methacrylate) spheres with a diameter of 30 μm and four different film thicknesses (270 nm, 150 nm, 100 nm, and 60 nm) were investigated. By multiplying the experimental results with geometrical correction factors obtained using finite element models, the resistivities of the thin films were estimated for the four thicknesses. These were higher than the resistivity of bulk silver.

SilE is an intrinsically disordered periplasmic "molecular sponge" involved in bacterial silver resistance.

SummaryAg+ resistance was initially found on the Salmonella enetrica serovar Typhimurium multi‐resistance plasmid pMG101 from burns patients in 1975. The putative model of Ag+ resistance, encoded by the sil operon from pMG101, involves export of Ag+ via an ATPase (SilP), an effluxer complex (SilCFBA) and a periplasmic chaperon of Ag+ (SilE). SilE is predicted to be intrinsically disordered. We tested this hypothesis using structural and biophysical studies and show that SilE is an intrinsically disordered protein in its free apo‐form but folds to a compact structure upon optimal binding to six Ag+ ions in its holo‐form. Sequence analyses and site‐directed mutagenesis established the importance of histidine and methionine containing motifs for Ag+‐binding, and identified a nucleation core that initiates Ag+‐mediated folding of SilE. We conclude that SilE is a molecular sponge for absorbing metal ions.

Contact Resistance and Metallurgical Connections Between Silver Coated Polymer Particles in Isotropic Conductive Adhesives

Recently, there has been an increasing interest in silver thin film coated polymer spheres as conductive fillers in isotropic conductive adhesives (ICAs). Such ICAs yield resistivities similar to conventional silver flake based ICAs while requiring only a fraction of the silver content. In this work, effects of the nanostructure of silver thin films on inter-particle contact resistance were investigated. The electrical resistivity of ICAs with similar particle content was shown to decrease with increasing coating thickness. Scanning electron micrographs of ion milled cross-sections revealed that the silver coatings formed continuous metallurgical connections at the contacts between the filler particles after adhesive curing at 150°C. The electrical resistivity decreased for all samples after environmental treatment for 3 weeks at 85°C/85% relative humidity. It was concluded that after the metallurgical connections formed, the bulk resistance of these ICAs were no longer dominated by the contact resistance, but by the geometry and nanostructure of the silver coatings. A figure of merit (FoM) was defined based on the ratio between bulk silver resistivity and the ICA resistivity, and this showed that although the resistivity was lowest in the ICAs containing the most silver, the volume of silver was more effectively used in the ICAs with intermediate silver contents. This was attributed to a size effect due to smaller grains in the thickest coating.

Evolution of a Heavy Metal Homeostasis/Resistance Island Reflects Increasing Copper Stress in Enterobacteria.

Copper homeostasis in bacteria is challenged by periodic elevation of copper levels in the environment, arising from both natural sources and human inputs. Several mechanisms have evolved to efflux copper from bacterial cells, including the cus (copper sensing copper efflux system), and pco (plasmid-borne copper resistance system) systems. The genes belonging to these two systems can be physically clustered in a Copper Homeostasis and Silver Resistance Island (CHASRI) on both plasmids and chromosomes in Enterobacteria. Increasing use of copper in agricultural and industrial applications raises questions about the role of human activity in the evolution of novel copper resistance mechanisms. Here we present evidence that CHASRI emerged and diversified in response to copper deposition across aerobic and anaerobic environments. An analysis of diversification rates and a molecular clock model suggest that CHASRI experienced repeated episodes of elevated diversification that could correspond to peaks in human copper production. Phylogenetic analyses suggest that CHASRI originated in a relative of Enterobacter cloacae as the ultimate product of sequential assembly of several pre-existing two-gene modules. Once assembled, CHASRI dispersed via horizontal gene transfer within Enterobacteriaceae and also to certain members of Shewanellaceae, where the original pco module was replaced by a divergent pco homolog. Analyses of copper stress mitigation suggest that CHASRI confers increased resistance aerobically, anaerobically, and during shifts between aerobic and anaerobic environments, which could explain its persistence in facultative anaerobes and emergent enteric pathogens.

Preventing Implant-Associated Infections by Silver Coating.

Implant-associated infections (IAIs) are a dreaded complication mainly caused by biofilm-forming staphylococci. Implant surfaces preventing microbial colonization would be desirable. We examined the preventive effect of a silver-coated titanium-aluminum-niobium (TiAlNb) alloy. The surface elicited a strong, inoculum-dependent activity againstStaphylococcus epidermidisandStaphylococcus aureusin an agar inhibition assay. Gamma sterilization and alcohol disinfection did not alter the effect. In a tissue cage mouse model, silver coating of TiAlNb cages prevented perioperative infections in an inoculum-dependent manner and led to a 100% prevention rate after challenge with 2 × 10(6)CFU ofS. epidermidisper cage. InS. aureusinfections, silver coating had only limited effect. Similarly, daptomycin or vancomycin prophylaxis alone did not preventS. aureusinfections. However, silver coating combined with daptomycin or vancomycin prophylaxis thwarted methicillin-resistantS. aureusinfections at a prevention rate of 100% or 33%, respectively. Moreover, silver release from the surface was independent of infection and occurred rapidly after implantation. On day 2, a peak of 82 μg Ag/ml was reached in the cage fluid, corresponding to almost 6× the MIC of the staphylococci. Cytotoxicity toward leukocytes in the cage was low and temporary. Surrounding tissue did not reveal histological signs of silver toxicity.In vitro, no emergence of silver resistance was observed in several clinical strains of staphylococci upon serial subinhibitory silver exposures. In conclusion, our data demonstrate that silver-coated TiAlNb is potent for prevention of IAIs and thus can be considered for clinical application.

RRR and thermal conductivity of Ag and Ag-0.2 wt.%Mg alloy in Ag/Bi-2212 wires

Residual resistivity ratio (RRR) and thermal conductivity of metal matrix in metal/superconductor composite wires are important parameters for designing superconducting magnets. However, the resistivity of silver in reacted Ag/Bi-2212 wires has yet to be determined over temperature range from 4.2 K to 80 K because Bi-2212 filaments have a critical transition temperature Tc of ∼ 80 K, and because it is unknown whether the RRR of Ag/Bi-2212 degrades with Cu diffusing from Bi-2212 filaments into silver sheathes at elevated temperatures and to what degree it varies with heat treatment. We measured the resistivity of stand-alone Ag and AgMg (Ag-0.2 wt.% Mg) wires as well as the resistivity of Ag and Ag-0.2 wt.% Mg in Ag/Bi- 2212 round wires reacted in 1 bar oxygen at 890 °C for 1, 8, 24 and 48 hours and quickly cooled to room temperature. The heat treatment was designed to reduce the critical current Ic of Bi- 2212 wires to nearly zero while allowing Cu loss to fully manifest itself. We determined that pure silver exhibits a RRR of ∼ 220 while the oxide-dispersion strengthened Ag-Mg exhibits a RRR of ∼ 5 in stand-alone samples. A surprising result is that the RRR of silver in the composite round wires doesn't degrade with extended time at 890 °C for up to 48 hours. This surprising result may be explained by our observation that the Cu that diffuses into silver tends to form Cu2O precipitates in oxidizing atmosphere, instead of forming Ag-Cu solution alloy. We also measured the thermal conductivity and the magneto-resistivity of pure Ag and Ag-0.2 wt. % Mg from 4.2 K to 300 K in magnetic fields up to 14.8 T and summarized them using a Kohler plot.

New Evidence Silver Resistance Is Emerging

New Evidence Silver Resistance Is Emerging

Unprecedented Silver Resistance in Clinically Isolated Enterobacteriaceae: Major Implications for Burn and Wound Management.

Increased utilization of inorganic silver as an adjunctive to many medical devices has raised concerns of emergent silver resistance in clinical bacteria. Although the molecular basis for silver resistance has been previously characterized, to date, significant phenotypic expression of these genes in clinical settings is yet to be observed. Here, we identified the first strains of clinical bacteria expressing silver resistance at a level that could significantly impact wound care and the use of silver-based dressings. Screening of 859 clinical isolates confirmed 31 harbored at least 1 silver resistance gene. Despite the presence of these genes, MIC testing revealed most of the bacteria displayed little or no increase in resistance to ionic silver (200 to 300 μM Ag(+)). However, 2 isolates (Klebsiella pneumonia and Enterobacter cloacae) were capable of robust growth at exceedingly high silver concentrations, with MIC values reaching 5,500 μM Ag(+). DNA sequencing of these two strains revealed the presence of genes homologous to known genetic determinants of heavy metal resistance. Darkening of the bacteria's pigment was observed after exposure to high silver concentrations. Scanning electron microscopy images showed the presence of silver nanoparticles embedded in the extracellular polymeric substance of both isolates. This finding suggested that the isolates may neutralize ionic silver via reduction to elemental silver. Antimicrobial testing revealed both organisms to be completely resistant to many commercially available silver-impregnated burn and wound dressings. Taken together, these findings provide the first evidence of clinical bacteria capable of expressing silver resistance at levels that could significantly impact wound management.

Synthesis and characterization of silver nanoparticles doped reduced graphene oxide

Colloidal silver is used to reduce resistance of reduced graphene oxide (GO). The silver nanoparticles are dispersed on the surface of the GO and intercalated into the layers. The silver doped GO can also form a flexible free standing film with the addition of a polymer. Electrochemical properties of the samples were investigated and find that the pseudo capacitive properties increase in the silver doped samples, resulting in an increase of the capacitance. The resistance of silver doped sample decreases dramatically as well. It, therefore, could be a good candidative electrode for energy storage devices and electrochemical sensors.

Rapid evolution of silver nanoparticle resistance in Escherichia coli.

The recent exponential increase in the use of engineered nanoparticles (eNPs) means both greater intentional and unintentional exposure of eNPs to microbes. Intentional use includes the use of eNPs as biocides. Unintentional exposure results from the fact that eNPs are included in a variety of commercial products (paints, sunscreens, cosmetics). Many of these eNPs are composed of heavy metals or metal oxides such as silver, gold, zinc, titanium dioxide, and zinc oxide. It is thought that since metallic/metallic oxide NPs impact so many aspects of bacterial physiology that it will difficult for bacteria to evolve resistance to them. This study utilized laboratory experimental evolution to evolve silver nanoparticle (AgNP) resistance in the bacterium Escherichia coli (K-12 MG1655), a bacterium that does not harbor any known silver resistance elements. After 225 generations of exposure to the AgNP environment, the treatment populations demonstrated greater fitness vs. control strains as measured by optical density (OD) and colony forming units (CFU) in the presence of varying concentrations of 10 nm citrate-coated silver nanoparticles (AgNP) or silver nitrate (AgNO3). Genomic analysis shows that changes associated with AgNP resistance were already accumulating within the treatment populations by generation 100, and by generation 200 three mutations had swept to high frequency in the AgNP resistance stocks. This study indicates that despite previous claims to the contrary bacteria can easily evolve resistance to AgNPs, and this occurs by relatively simple genomic changes. These results indicate that care should be taken with regards to the use of eNPs as biocides as well as with regards to unintentional exposure of microbial communities to eNPs in waste products.

Silver resistance in Gram-negative bacteria: a dissection of endogenous and exogenous mechanisms.

ObjectivesTo gain a more detailed understanding of endogenous (mutational) and exogenous (horizontally acquired) resistance to silver in Gram-negative pathogens, with an emphasis on clarifying the genetic bases for resistance.MethodsA suite of microbiological and molecular genetic techniques was employed to select and characterize endogenous and exogenous silver resistance in several Gram-negative species.ResultsIn Escherichia coli, endogenous resistance arose after 6 days of exposure to silver, a consequence of two point mutations that were both necessary and sufficient for the phenotype. These mutations, in ompR and cusS, respectively conferred loss of the OmpC/F porins and derepression of the CusCFBA efflux transporter, both phenotypic changes previously linked to reduced intracellular accumulation of silver. Exogenous resistance involved derepression of the SilCFBA efflux transporter as a consequence of mutation in silS, but was additionally contingent on expression of the periplasmic silver-sequestration protein SilE. Silver resistance could be selected at high frequency (>10−9) from Enterobacteriaceae lacking OmpC/F porins or harbouring the sil operon and both endogenous and exogenous resistance were associated with modest fitness costs in vitro.ConclusionsBoth endogenous and exogenous silver resistance are dependent on the derepressed expression of closely related efflux transporters and are therefore mechanistically similar phenotypes. The ease with which silver resistance can become selected in some bacterial pathogens in vitro suggests that there would be benefit in improved surveillance for silver-resistant isolates in the clinic, along with greater control over use of silver-containing products, in order to best preserve the clinical utility of silver.

Use of bioreporters and deletion mutants reveals ionic silver and ROS to be equally important in silver nanotoxicity

The mechanism of antibacterial action of silver nanoparticles (AgNp) was investigated by employing a combination of microbiology and geochemical approaches to contribute to the realistic assessment of nanotoxicity. Our studies showed that suspending AgNp in media with different levels of chloride relevant to environmental conditions produced low levels of ionic silver thereby suggesting that dissolution of silver ions from nanoparticulate surface could not be the sole mechanism of toxicity. An Escherichia coli based bioreporter strain responsive to silver ions together with mutant strains of E. coli lacking specific protective systems were tested against AgNp. Deletion mutants lacking silver ion efflux systems and resistance mechanisms against oxidative stress showed an increased sensitivity to AgNp. However, the bioreporter did not respond to silver nanoparticles. Our results suggest that oxidative stress is a major toxicity mechanism and that this is at least partially associated with ionic silver, but that bulk dissolution of silver into the medium is not sufficient to account for the observed effects. Chloride ions do not appear to offer significant protection, indicating that chloride in receiving waters will not necessarily protect environmental bacteria from the toxic effects of nanoparticles in effluents.

Preparation of Ag/Cu/Ti Nanofluids by Spark Discharge System and Its Control Parameters Study

This study selected silver, copper, and titanium as the research objects to explore the relationship between nanofluids properties and electrical discharge machining (EDM) processes. Regarding the products, UV-visible spectroscopy (UV-Vis) was applied to measure the concentration distribution of nanofluids; zeta-size analysis is applied for measuring nanometal particles’ Zeta-Potential and the size distribution of metallic particles in the fluid. Finally, various instruments, including scanning electron microscope (SEM), were applied to observe the shape, size, and composition ratio of metal particles after processing. According to the experimental results, the control of the discharge pulse time, in addition to affecting the concentration of metallic liquid and temperature in the process, affects the size of the metal particles after the process. As the resistivity of silver and copper is very low, at about15×10-9 Ω·m, ifTONis set to between 10~50 μs, good preparation efficiency can be obtained. The resistivity of titanium is420×10-9 Ω·m, which is much larger than that of silver or copper. Hence,TONshould be set to approximately 100 μs to achieve a good discharge success rate.

Atomic Force Microscope Spectroscopy: Progress toward Antibiotic Resistance and Biofilm Studies

The emerging field of live cell nanoscopy and force spectroscopy could revolutionize the way biologists explore the living cell at a molecular resolution. Atomic Force Microscope (AFM) and force spectroscopy analysis have been used to directly measure reversible physiochemical and specific binding interactions between cells. Stickiness is important biofilm formation stage that could also be measured at nN level. A significant source of foodborne illness results from biofilms. These are caused by microorganisms that attach to surfaces and grow as highly organized multicellular communities. This study examines the impact of silver resistance on bacterial adhesion and its viscoelastic formation. We present the first set of data that evaluates the elastic deformation of a bacterial cell surface upon evolution of silver resistance in E. coli MG1655 using AFM compared to controls in 200 generations. The adhesion stickiness and stiffness mean of the treated (evolved Ag resistant) and non-treated samples were tested at nN level. The evolved samples had a significantly (P<0.05) higher stickiness ratio and value (from 0.01± 0.04 nN to 0.06± 0.02 nN) compared to the controls (non-resistant) strains (from0.01± 0.02 nN to0.04± 0.02 nN). The highest difference of adhesion force happened on Generation 100. According to images of the bacteria in different generations, we can see some major phenotype changes on the appearance at generation 100. The MIC data for the non-evolved strains of E. coli MG 1655 through 200 generations were also significantly lower 38.58± 10.09 mg/l compare to evolved strains 272.25 ± 153.94 mg/l (p<0.01). The experiment demonstrates important features of phenotype modulation resulting from the evolution of Ag resistance that will be further studied by this group.

A long-term study examining the antibacterial effectiveness of Agion silver zeolite technology on door handles within a college campus.

Laboratory studies have shown that small concentrations of silver are effective at inhibiting the growth micro-organisms through the disruption of important cell structures and processes. The additional ability to incorporate silver into surfaces has increased the usage of silver in the medical field and expanded its use into the consumer market. To understand the impact of increased silver-containing antimicrobial use, it is important to determine whether silver-based consumer goods are effective at reducing bacterial populations. Our study examined the antibacterial effectiveness of Agion silver zeolite technology applied to 25 silver- and control-coated door handles across a college campus. Door handles were sampled for 6 week periods in both the fall and spring semester, and bacteria were cultured and enumerated on tryptic soy agar (TSA), MacConkey agar (MAC) and mannitol salt agar (MSA). A significant difference was observed between the bacterial populations isolated from silver- and control-coated door handles after 3 years. However, bacteria were consistently isolated from silver-coated door handles suggesting that the silver zeolite was only effective against a portion of the bacterial populations, and further studies are necessary to determine the identities of the isolated bacteria and the prevalence of silver resistance.

Lysozyme-coated silver nanoparticles for differentiating bacterial strains on the basis of antibacterial activity

Lysozyme, an antibacterial enzyme, was used as a stabilizing ligand for the synthesis of fairly uniform silver nanoparticles adopting various strategies. The synthesized particles were characterized using UV-visible spectroscopy, FTIR, dynamic light scattering (DLS), and TEM to observe their morphology and surface chemistry. The silver nanoparticles were evaluated for their antimicrobial activity against several bacterial species and various bacterial strains within the same species. The cationic silver nanoparticles were found to be more effective against Pseudomonas aeruginosa 3 compared to other bacterial species/strains investigated. Some of the bacterial strains of the same species showed variable antibacterial activity. The difference in antimicrobial activity of these particles has led to the conclusion that antimicrobial products formed from silver nanoparticles may not be equally effective against all the bacteria. This difference in the antibacterial activity of silver nanoparticles for different bacterial strains from the same species may be due to the genome islands that are acquired through horizontal gene transfer (HGT). These genome islands are expected to possess some genes that may encode enzymes to resist the antimicrobial activity of silver nanoparticles. These silver nanoparticles may thus also be used to differentiate some bacterial strains within the same species due to variable silver resistance of these variants, which may not possible by simple biochemical tests.

Study on the Heat Transfer Characteristics of Warm Needle Materials in Korean Medicine

Objectives : Warm needling is the method combining the effects of acupuncture and moxibustion. In modern clinics, warm needling is only affected by treatment time or frequency. We need to study the physical characteristics of warm needles currently used in clinics in order to develop more efficient and economical treatments. Methods : We collected various warm needle specimens and analyzed chemical constitutions, measured heat transfer velocity, micro Vicker`s hardness and specific resistance. We studied the relationship between heat transfer velocity and micro Vicker`s hardness as well as that between heat transfer velocity and specific resistance. Results : The heat transfer velocity of the Silver_HL was 3.3 mm/sec, of the Au alloy group was 1.3~2.6 mm/sec, and Silver_IN, Silver_ZK and SS groups was less than 0.3 mm/sec. We therefore concluded that the needle composed of Ag has the best heat transfer velocity. In the Micro Vicker`s hardness test, Vicker`s hardness of the Au alloy group was 159~170 Hv, of Silver_HL was 181 Hv, and of the Silver_IN, Silver_ZK, SS group was 450.8~519 Hv. In the Silver_IN, Silver_ZK and SS groups, hardness was inversely proportial to thermal conductivity. In the specific resistance test, the specific resistance of Silver_HL was the lowest, that of the Au alloy group was the second lowest, and that of the Silver_IN, Silver_ZK, SS groups were the highest. Conclusions : We concluded that the needle composed of Ag has the best heat transfer velocity, highest electric conductivity and thermal conductivity, therefore the needle composed of Ag is suitable for warm needling.

Silver resistance genes are overrepresented among Escherichia coli isolates with CTX-M production.

Members of the Enterobacteriaceae with extended-spectrum beta-lactamases (ESBLs) of the CTX-M type have disseminated rapidly in recent years and have become a threat to public health. In parallel with the CTX-M type expansion, the consumption and widespread use of silver-containing products has increased. To determine the carriage rates of silver resistance genes in different Escherichia coli populations, the presence of three silver resistance genes (silE, silP, and silS) and genes encoding CTX-M-, TEM-, and SHV-type enzymes were explored in E. coli isolates of human (n = 105) and avian (n = 111) origin. The antibiotic profiles were also determined. Isolates harboring CTX-M genes were further characterized, and phenotypic silver resistance was examined. The silE gene was present in 13 of the isolates. All of them were of human origin. Eleven of these isolates harbored ESBLs of the CTX-M type (P = 0.007), and eight of them were typed as CTX-M-15 and three as CTX-M-14. None of the silE-positive isolates was related to the O25b-ST131 clone, but 10 out of 13 belonged to the ST10 or ST58 complexes. Phenotypic silver resistance (silver nitrate MIC > 512 mg/liter) was observed after silver exposure in 12 of them, and a concomitant reduced susceptibility to piperacillin-tazobactam developed in three. In conclusion, 12% of the human E. coli isolates but none of the avian isolates harbored silver resistance genes. This indicates another route for or level of silver exposure for humans than that caused by common environmental contamination. Since silE-positive isolates were significantly more often found in CTX-M-positive isolates, it is possible that silver may exert a selective pressure on CTX-M-producing E. coli isolates.