Superior infectivity for mosquito vectors contributes to competitive displacement among strains of dengue virus

<p>Abstract</p> <p>Background</p> <p>Competitive displacement of a weakly virulent pathogen strain by a more virulent strain is one route to disease emergence. However the mechanisms by which pathogens compete for access to hosts are poorly understood. Among vector-borne pathogens, variation in the ability to infect vectors may effect displacement. The current study focused on competitive displacement in dengue virus serotype 3 (DENV3), a mosquito-borne pathogen of humans. In Sri Lanka in the 1980's, a native DENV3 strain associated with relatively mild dengue disease was displaced by an invasive DENV3 strain associated with the most severe disease manifestations, dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS), resulting in an outbreak of DHF/DSS. Here we tested the hypothesis that differences between the invasive and native strain in their infectivity for <it>Aedes aegypti </it>mosquitoes, the primary vector of DENV, contributed to the competitive success of the invasive strain</p> <p>Results</p> <p>To be transmitted by a mosquito, DENV must infect and replicate in the midgut, disseminate into the hemocoel, infect the salivary glands, and be released into the saliva. The ability of the native and invasive DENV3 strains to complete the first three steps of this process in <it>Aedes aegypti </it>mosquitoes was measured <it>in vivo</it>. The invasive strain infected a similar proportion of mosquitoes as the native strain but replicated to significantly higher titers in the midgut and disseminated with significantly greater efficiency than the native strain. In contrast, the native and invasive strain showed no significant difference in replication in cultured mosquito, monkey or human cells.</p> <p>Conclusion</p> <p>The invasive DENV3 strain infects and disseminates in <it>Ae. aegypti </it>more efficiently than the displaced native DENV3 strain, suggesting that the invasive strain is transmitted more efficiently. Replication in cultured cells did not adequately characterize the known phenotypic differences between native and invasive DENV3 strains. Infection dynamics within the vector may have a significant impact on the spread and replacement of dengue virus lineages.</p

Metal ions and graphene-based compounds as alternative treatment options for burn wounds infected by antibiotic-resistant Pseudomonas aeruginosa.

Burn infections caused by Pseudomonas aeruginosa pose a major complication in wound healing. This study aimed to determine the antimicrobial effect of metal ions, graphene (Gr), and graphene oxide (GO), individually and in combination, against the planktonic and biofilm states of two antimicrobially resistant clinical strains of P. aeruginosa each with different antibiotic resistance profiles. Minimum inhibitory, minimum bactericidal, and fractional inhibitory concentrations were performed to determine the efficacy of the metal ions and graphene composites individually and their synergy in combination. Crystal violet biofilm and XTT assays measured the biofilm inhibition and metabolic activity, respectively. Molybdenum, platinum, tin, gold, and palladium ions exhibited the greatest antimicrobial activity (MIC = 7.8-26.0 mg/L), whilst GO and Gr demonstrated moderate-to-no effect against the planktonic bacterial cells, irrespective of their antibiograms. Biofilms were inhibited by zinc, palladium, silver, and graphene. In combination, silver-graphene and molybdenum-graphene inhibited both the planktonic and biofilm forms of the bacteria making them potential candidates for development into topical antimicrobials for burns patients infected with antibiotic-resistant P. aeruginosa

Graphene derivatives potentiate the activity of antibiotics against Enterococcus faecium, Klebsiella pneumoniae and Escherichia coli

Antibiotic resistance in bacteria is developing at a faster rate than new antibiotics can be discovered. This study investigated the antimicrobial activity of several carbon-based derivative compounds alone and in combination with clinically relevant antibiotics against key ESKAPE pathogens Enterococcus faecium, Klebsiella pneumoniae and Escherichia coli. Three compounds, graphite, graphene and graphene oxide, in conjunction with ciprofloxacin (CIP), chloramphenicol (CHL) and piperacillin/tazobactam (TZP) were examined using fractional inhibitory concentration (FIC) testing. CIP combined with graphene demonstrated additive antimicrobial activity against E. faecium compared to individual application. Furthermore, CIP supplemented with graphene, graphene oxide or graphite showed additive activity with ∑FIC values of 1.0 against K. pneumoniae, whereas only TZP showed ∑FIC value

Non-chlorine detergent formulations as an alternative for unpasteurised milk removal from stainless steel surfaces

Hygiene is a major concern in the dairy industry, and detergents based on hypochlorite have commonly been utilised for cleaning-in-place (CIP) regimes. However, due to concerns about chlorate residues entering the milk processing chain, new detergent alternatives that are free of chlorate sources are required. Two new formulations were developed based on ethylenediaminetetraacetic acid (EDTA) and wetting agents. Stainless steel surfaces were fouled with milk and cleaned once or 10 times using water, a caustic-EDTA product, a sodium hypochlorite product, an acid, or new cleaning formulations (Product A and Product B). The results demonstrated that the use of acid did not result in successful cleaning. The two new products performed well, with Product B performing equivalently during cleaning compared with the established hypochlorite or caustic-EDTA products. Product A exhibited better cleanliness than the other detergents tested. When allergen removal was considered, residual material was found to be retained on the surfaces, regardless of the cleaning type used. This study suggests that the new product formulations may be used to replace hypochlorite-based detergents to increase the hygienic status of a surface

Photodynamic antimicrobial chemotherapy coupled with the use of the photosensitizers methylene blue and temoporfin as a potential novel treatment for Staphylococcus aureus in burn infections

Photodynamic antimicrobial chemotherapy (PACT) is a novel alternative antimicrobial therapy that elicits a broad mechanism of action and therefore has a low probability of generating resistance. Such properties make PACT ideally suited for utilization in localized applications such as burn wounds. The aim of this study was to determine the antimicrobial activity of MB and temoporfin against both a S. aureus isolate and a P. aeruginosa isolate in light (640 nm) and dark conditions at a range of time points (0–20 min). A Staphylococcus aureus isolate and a Pseudomonas aeruginosa isolate were treated in vitro with methylene blue (MB) and temoporfin under different conditions following exposure to light at 640 nm and in no-light (dark) conditions. Bacterial cell viability [colony-forming units (c.f.u.) ml−1] was then calculated. Against P. aeruginosa , when MB was used as the photosensitizer, no phototoxic effect was observed in either light or dark conditions. After treatment with temoporfin, a reduction of less than one log (7.00×107 c.f.u. ml−1) was observed in the light after 20 min of exposure. However, temoporfin completely eradicated S. aureus in both light and dark conditions after 1 min (where a seven log reduction in c.f.u. ml−1 was observed). Methylene blue resulted in a loss of S. aureus viability, with a two log reduction in bacterial viability (c.f.u. ml−1) reported in both light and dark conditions after 20 min exposure time. Temoporfin demonstrated greater antimicrobial efficacy than MB against both the S. aureus and P. aeruginosa isolates tested. At 12.5 µM temoporfin resulted in complete eradication of S. aureus . In light of this study, further research into the validity of PACT, coupled with the photosensitizers (such as temoporfin), should be conducted in order to potentially develop alternative antimicrobial treatment regimes for burn wounds.</jats:p

The Influence of Surface Topography and Wettability on Escherichia coli Removal from Polymeric Materials in the Presence of a Blood Conditioning Film

The reduction of biofouling and the reduction of cross-contamination in the food industry are important aspects of safety management systems. Polymeric surfaces are used extensively throughout the food production industry and therefore ensuring that effective cleaning regimes are conducted is vital. Throughout this study, the influence of the surface characteristics of three different polymeric surfaces, polytetrafluoroethylene (PTFE), poly(methyl methacrylate) (PMMA) and polyethylene terephthalate (PET), on the removal of Escherichia coli using a wipe clean method utilising 3% sodium hypochlorite was determined. The PTFE surfaces were the roughest and demonstrated the least wettable surface (118.8°), followed by the PMMA (75.2°) and PET surfaces (53.9°). Following cleaning with a 3% sodium hypochlorite solution, bacteria were completely removed from the PTFE surfaces, whilst the PMMA and PET surfaces still had high numbers of bacteria recovered (1.2 × 107 CFU/mL and 6.3 × 107 CFU/mL, respectively). When bacterial suspensions were applied to the surfaces in the presence of a blood conditioning film, cleaning with sodium hypochlorite demonstrated that no bacteria were recovered from the PMMA surface. However, on both the PTFE and PET surfaces, bacteria were recovered at lower concentrations (2.0 × 102 CFU/mL and 1.3 × 103 CFU/mL, respectively). ATP bioluminescence results demonstrated significantly different ATP concentrations on the surfaces when soiled (PTFE: 132 relative light units (RLU), PMMA: 80 RLU and PET: 99 RLU). Following cleaning, both in the presence and absence of a blood conditioning film, all the surfaces were considered clean, producing ATP concentrations in the range of 0–2 RLU. The results generated in this study demonstrated that the presence of a blood conditioning film significantly altered the removal of bacteria from the polymeric surfaces following a standard cleaning regime. Conditioning films which represent the environment where the surface is intended to be used should be a vital part of the test regime to ensure an effective disinfection process

Exploring the reactivity of distinct electron transfer sites at CVD grown monolayer graphene through the selective electrodeposition of MoO2 nanowires

The origin of electron transfer at Chemical Vapour Deposition (CVD) grown monolayer graphene using a polymer-free transfer methodology is explored through the selective electrodeposition of Molybdenum (di)oxide (MoO2). The electrochemical decoration of CVD monolayer graphene with MoO2 is shown to originate from the edge plane like- sites/defects. Edge plane decoration of MoO2 nanowires upon monolayer graphene is observed via electrochemical deposition over short time periods only (ca. −0.6 V for 1 second (vs. Ag/AgCl)). At more electrochemically negative potentials (ca. −1.0 V) or longer deposition times, a large MoO2 film is created/deposited on the graphene sheet, originating and expanding from the original nucleation points at edge plane like- sites/defects/wrinkles. Nanowire fabrication along the edge plane like- sites/defects of graphene is confirmed with Cyclic Voltammetry, Scanning Electron Microscopy (SEM), Atomic Force Microscopy (AFM) and Raman Spectroscopy. Monitoring the electrochemical response towards [Ru(NH3)6]3+/2+ and comparing the heterogeneous electron transfer (HET) kinetics at CVD grown monolayer graphene prior and post nanowire fabrication reveals key understandings into the fundamental electrochemical properties of carbon materials. The HET kinetics (k0obs) at MoO2 nanowire decorated monolayer graphene sheets, when edge plane like- sites/defects have been coated/blocked with MoO2, are significantly reduced in comparison to the unmodified graphene alternative. Interestingly, MoO2 nucleation originates on the edge plane like- sites/defects of the graphene sheets, where the basal plane sites remain unaltered until the available edge plane like- sites/defects have been fully utilised; after which MoO2 deposition propagates towards and onto the basal planes, eventually covering the entire surface of the monolayer graphene surface. In such instances, there is no longer an observable electrochemical response. This work demonstrates the distinct electron transfer properties of edge and basal plane sites on CVD grown monolayer graphene, inferring favourable electrochemical reactivity at edge plane like- sites/defects and clarifying the origin of graphene electro-activity

A trade-off in replication in mosquito versus mammalian systems conferred by a point mutation in the NS4B protein of dengue virus type 4

AbstractAn acceptable live-attenuated dengue virus vaccine candidate should have low potential for transmission by mosquitoes. We have identified and characterized a mutation in dengue virus type 4 (DEN4) that decreases the ability of the virus to infect mosquitoes. A panel of 1248 mutagenized virus clones generated previously by chemical mutagenesis was screened for decreased replication in mosquito C6/36 cells but efficient replication in simian Vero cells. One virus met these criteria and contained a single coding mutation: a C-to-U mutation at nucleotide 7129 resulting in a Pro-to-Leu change in amino acid 101 of the nonstructural 4B gene (NS4B P101L). This mutation results in decreased replication in C6/36 cells relative to wild-type DEN4, decreased infectivity for mosquitoes, enhanced replication in Vero and human HuH-7 cells, and enhanced replication in SCID mice implanted with HuH-7 cells (SCID-HuH-7 mice). A recombinant DEN4 virus (rDEN4) bearing this mutation exhibited the same set of phenotypes. Addition of the NS4B P101L mutation to rDEN4 bearing a 30 nucleotide deletion (Δ30) decreased the ability of the double-mutant virus to infect mosquitoes but increased its ability to replicate in SCID-HuH-7 mice. Although the NS4B P101L mutation decreases infectivity of DEN4 for mosquitoes, its ability to enhance replication in SCID-HuH-7 mice suggests that it might not be advantageous to include this specific mutation in an rDEN4 vaccine. The opposing effects of the NS4B P101L mutation in mosquito and vertebrate systems suggest that the NS4B protein is involved in maintaining the balance between efficient replication in the mosquito vector and the human host

Evaluation of a microfluidic atmospheric-pressure plasma reactor for water treatment

A dielectric barrier discharge (DBD) microfluidic plasma reactor, operated at atmospheric pressure, was studied for its potential to treat organic contaminants in water. The proposed microfluidic plasma reactor (MPR) allows in situ production of plasma in a continuous flow, operated under atmospheric pressure, for plasma-based water treatment. The MPR operates with glass as the dielectric barrier, where plasma is generated in the continuous gas flow stream of a gas-liquid two-phase annular flow regime in the microchannels. The microchannels have dimensions of 100 µm depth, 250 µm width and the plasma is generated in an approximately 21 cm length of microchannel arranged in a serpentine pattern. Methylene blue (MB) in solution was used as a model organic to investigate its degradation by plasma generated in the microchannels. The influence of discharge time, residence time and gas sources, i.e. air, argon and oxygen, on MB degradation was studied. The percentage degradation increased with lower liquid flow rates, with maximum degradation of MB achieved at a liquid flow rate of 35 µL/min and inlet gas pressure of 1 bar using oxygen as the working gas. Liquid chromatography/mass spectrometry analysis of the MB solution after treatment suggests degradation through fragmentation of MB. It is intended that the device will be used as proof of concept to introduce plasma technology as an advanced oxidation process for water treatment, with the potential to achieve total mineralization of dissolved organic materials and microbial inactivation, replacing water treatment chemicals and consumables

Interfacial surface properties of compression moulded hydrolysed Polyvinyl Acetate (PVAc) using different release materials

Understanding the effect surface production has on polymer properties is important in the design of advanced materials. The aim of this study was to determine how the moulding process affected the rate of hydrolysis and the topography, chemistry and physicochemistry of PVAc moulded, hydrolysed surfaces. Three different mould surface materials were used to produce compression moulded PVAc sheets which were treated with aqueous NaOH at a range of concentrations. The Textile moulded sheet demonstrated the best hydrolysis results. The topography of the moulded sheets was transferred to the surfaces and the Kapton release sheet was visually smooth at lower magnification and demonstrated some pitting at higher magnification. The Teflon surface had features transferred from the coated stainless steel at lower magnifications and linear features at higher magnifications and the textile surface had a wrinkled appearance and irregularly spaced peaks. The release sheet used to mould the PVAc surfaces, affected the physicochemical parameters. The Kapton moulded surface demonstrated the most polar attributes and the Teflon surface the most dispersive. It was clear that the selection of the mould material had an influence on surface properties and hydrolysis of moulded PVAc. Such information is important for engineering design in industrial processes