Search results for: MS2 virus inactivation
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 733

Search results for: MS2 virus inactivation

733 Household Low Temperature MS2 (ATCC15597-B1) Virus Inactivation Using a Hot Bubble Column Evaporator

Authors: Adrian Garrido Sanchis, Richard Pashley

Abstract:

The MS2 (ATCC15597-B1) virus was used as a surrogate to estimate the inactivation rates for enteric viruses when using a hot air bubble column evaporator (HBCE) system in the treatment of household wastewater. In this study, we have combined MS2 virus surface charging properties with thermal inactivation rates, using an improved double layer plaque assay technique, in order to assess the efficiency of the HBCE process for virus removal in water. When bubbling a continuous flow of dry air, at 200°C, only heats the aqueous solution in the bubble column to about 50°C. Viruses are not inactivated by this solution temperature, as confirmed separately from water bath heating experiments. Hence, the efficiency of the HBCE process for virus removal in water appeared to be caused entirely by collisions between the hot air bubbles and the virus organisms. This new energy efficient treatment for water reuse applications can reduce the thermal energy required to only 25% (about 113.7 kJ/L) of that required for boiling (about 450 kJ/L).

Keywords: MS2 virus inactivation, water reuse, hot bubble column evaporator, water treatment

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732 Inactivation Kinetics of DNA and RNA Viruses by Ozone-Air Mixture in a Flow Mixer

Authors: Nikolai Nosik, Vladislav Podmasterjev, Nina Kondrashina, Marina Chataeva, Olga Lobach, Dmitry Noosik, Sergei Razumovskii

Abstract:

Virucidal activity of ozone is well known: dissolved in water it kill viruses very fast. The virucidal capacity of ozone in ozone-air mixture is less known. The goal of the study was to investigate the virucidal potentials of the ozone–air mixture and kinetics of virus inactivation. Materials and methods. Ozone (O3 ) was generated from oxygen with ozonizer ( 1.0 – 75.0 mg\l). The ozone concentration was determined by the spectrophotometric methods. Virus contaminated samples were placed into the flowing reactor. Viruses: poliovirus type 1, vaccine strain (Sabin) and adenovirus, type 5, were obtained from the State virus collection. Titrations of viruses were carried out in appropriate cell cultures. CxT value ( mg\l x min) was calculated. Results. Metallic, polycarbonic and fiber “Kevlar” samples were contaminated with virus, dried and treated with ozone-air mixture in the flowing reactor. Kinetics of poliovirus inactivation: in 15 min at 5.0 mg\l -2.0 lg TCID50 inhibition , in 15 min at 10 mg\l – 2.5 lg TCID50 , 4.0 lg TCID50 inactivation of poliovirus was achieved after 75min at ozone concentration 20.0mg\l (99.99%). ( CxT = 75, 150 and 1500 mg\l x min on all three types of surfaces). It was found that the inactivation of poliovirus was more effective when the virus contaminated samples were wet (in 15 min at 20mg\l inhibition of virus in dry samples was 2.0 TCID50 , in wet samples – 4.0 TCID50). Adenovirus was less resistant to ozone treatment then poliovirus: 4.0 lg TCID50 inhibition was observed after 30 min of the treatment with ozone at 20mg\l ( CxT mg\l x min = 300 for adenovirus as for poliovirus it was 1500). Conclusion. It was found that ozone-air mixture inactivates viruses at rather high concentrations (compared to the reported effect of ozone dissolved in water). Despite of that there is a difference in the resistance to ozone action between viruses – poliovirus is more resistant then adenovirus-ozone-air mixture can be used for disinfection of large rooms. The maintaining of the virus-contaminated surfaces in wet condition allow to decrease the ozone load for virus inactivation.

Keywords: adenovirus, disinfection, ozone, poliovirus

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731 Diverse Sensitivity to Ultraviolet Radiation of DNA and RNA Viruses

Authors: Nickolay Nosik, Dmitry Nosik, Marina Bochkova, Nina Kondrashina, Olga Lobach

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The bactericidal effect of UV radiation is known for long time and widely used for inactivation of pathogens but for viruses it is not so uniform. Due to a wide variety of viruses their sensitivity to UV radiation is quite different and not quite predictable. The goal of the study was to determine the inactivation kinetics of UV radiation ( 254 nm) of the viruses of social importance (HIV), as well as test-viruses (poliovirus, adenovirus) used for the evaluation of the viral inactivation efficacy of germicides. Methods: DNA viruses- adenovirus, type 5; Herpes simplex virus (HSV), type 1, and RNA viruses–human immunodeficiency virus (HIV), type 1 and poliovirus, type 1 (Sabin strain) were obtained from State collection of viruses ( The D.I. Ivanovsky Institute of Virology). The source of UV radiation was a 15-watt low-pressure mercury vapor lamp (over 60% 254nm). The samples of 5cm2 were placed direct under the UV lamp flow (h-0.3m). Log reduction value was used as a marker for the rate of virus inactivation. Results: The data obtained indicate that poliovirus (one of the viruses most resistant to chemical germicides) and HSV are rather sensitive to UV radiation ( D90 =250-311 J/m2). Adenovirus is much more resistant to UV radiation (750 J/m2 ). The kinetics of adenovirus inactivation : 0 min- 5.0 lg TCID50, 10 min - 5,0, 15 min -4,0, 30 min – 3.5, 60 min – 1,0, 75 min -0,5 lg TCID50, 90 min –virus not detectable. HIV is most resistant to UV radiation among the studied viruses. It takes more than 4 hrs to inactivate the virus on the surface. D90 = 2000 J/m2 Conclusion: The results of the study show that there is no direct dependence between sensitivity to UV light and the size of the virion or presence\absence of the envelope of the virus. Poliovirus and adenovirus are small viruses (20-30nm poliovirus and 70-90nm adenovirus) and both are non-enveloped viruses but adenovirus 3-fold more resistant to UV radiation than poliovirus. It can be expected that viruses with more complicate structure, like Herpes virus (200nm) or HIV (80-100 nm), would be more sensitive to UV light. However, the very high resistance of HIV to UV radiation needs further investigation. The diverse resistance of the different viruses to UV radiation should be taken into the account when UV light is used to inactivate infectious viruses in hospitals and other public environments.

Keywords: HIV, HSV, inhibition of viruses, UV radiation

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730 Inactivation of Listeria innocua ATCC 33092 by Gas-Phase Plasma Treatment

Authors: Z. Herceg, V. Stulic, T. Vukusic, A. Rezek Jambrak

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High voltage electrical discharge plasmas are new nonthermal developing techniques used for water decontamination. To the full understanding of cell inactivation mechanisms, this study brings inactivation, recovery and cellular leakage of L. innocua cells before and after the treatment. Bacterial solution (200 mL) of L. innocua was treated in a glass reactor with a point-to-plate electrode configuration (high voltage electrode-titanium wire, was in the gas phase and grounded electrode was in the liquid phase). Argon was injected into the headspace of the reactor at the gas flow of 5 L/min. Frequency of 60, 90 and 120 Hz, time of 5 and 10 min, positive polarity and conductivity of media of 100 µS/cm were chosen to define listed parameters. With a longer treatment time inactivation was higher as well as the increase in cellular leakage. Despite total inactivation recovery of cells occurred probably because of a high leakage of proteins, compared to lower leakage of nucleic acids (DNA and RNA). In order to define mechanisms of inactivation further research is needed.

Keywords: Listeria innocua ATCC 33092, inactivation, gas phase plasma, cellular leakage, recovery of cells

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729 The Effect of Electrical Discharge Plasma on Inactivation of Escherichia Coli MG 1655 in Pure Culture

Authors: Zoran Herceg, Višnja Stulić, Anet Režek Jambrak, Tomislava Vukušić

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Electrical discharge plasma is a new non-thermal processing technique which is used for the inactivation of contaminating and hazardous microbes in liquids. Plasma is a source of different antimicrobial species including UV photons, charged particles, and reactive species such as superoxide, hydroxyl radicals, nitric oxide and ozone. Escherichia coli was studied as foodborne pathogen. The aim of this work was to examine inactivation effects of electrical discharge plasma treatment on the Escherichia coli MG 1655 in pure culture. Two types of plasma configuration and polarity were used. First configuration was with titanium wire as high voltage needle and another with medical stainless steel needle used to form bubbles in treated volume and titanium wire as high voltage needle. Model solution samples were inoculated with Escerichia coli MG 1655 and treated by electrical discharge plasma at treatment time of 5 and 10 min, and frequency of 60, 90 and 120 Hz. With the first configuration after 5 minutes of treatment at frequency of 120 Hz the inactivation rate was 1.3 log₁₀ reduction and after 10 minutes of treatment the inactivation rate was 3.0 log₁₀ reduction. At the frequency of 90 Hz after 10 minutes inactivation rate was 1.3 log₁₀ reduction. With the second configuration after 5 minutes of treatment at frequency of 120 Hz the inactivation rate was 1.2 log₁₀ reduction and after 10 minutes of treatment the inactivation rate was also 3.0 log₁₀ reduction. In this work it was also examined the formation of biofilm, nucleotide and protein leakage at 260/280 nm, before and after treatment and recuperation of treated samples. Further optimization of method is needed to understand mechanism of inactivation.

Keywords: electrical discharge plasma, escherichia coli MG 1655, inactivation, point-to-plate electrode configuration

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728 Numerical Simulation of Ultraviolet Disinfection in a Water Reactor

Authors: H. Shokouhmand, H. Sobhani, B. Sajadi, M. Degheh

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In recent years, experimental and numerical investigation of water UV reactors has increased significantly. The main drawback of experimental methods is confined and expensive survey of UV reactors features. In this study, a CFD model utilizing the eulerian-lagrangian framework is applied to analysis the disinfection performance of a closed conduit reactor which contains four UV lamps perpendicular to the flow. A discrete ordinates (DO) model was employed to evaluate the UV irradiance field. To investigate the importance of each of lamps on the inactivation performance, in addition to the reference model (with 4 bright lamps), several models with one or two bright lamps in various arrangements were considered. All results were reported in three inactivation kinetics. The results showed that the log inactivation of the two central bright lamps model was between 88-99 percent, close to the reference model results. Also, whatever the lamps are closer to the main flow region, they have more effect on microbial inactivation. The effect of some operational parameters such as water flow rate, inlet water temperature, and lamps power were also studied.

Keywords: Eulerian-Lagrangian framework, inactivation kinetics, log inactivation, water UV reactor

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727 Micropillar-Assisted Electric Field Enhancement for High-Efficiency Inactivation of Bacteria

Authors: Sanam Pudasaini, A. T. K. Perera, Ahmed Syed Shaheer Uddin, Sum Huan Ng, Chun Yang

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Development of high-efficiency and environment friendly bacterial inactivation methods is of great importance for preventing waterborne diseases which are one of the leading causes of death in the world. Traditional bacterial inactivation methods (e.g., ultraviolet radiation and chlorination) have several limitations such as longer treatment time, formation of toxic byproducts, bacterial regrowth, etc. Recently, an electroporation-based inactivation method was introduced as a substitute. Here, an electroporation-based continuous flow microfluidic device equipped with an array of micropillars is developed, and the device achieved high bacterial inactivation performance ( > 99.9%) within a short exposure time ( < 1 s). More than 99.9% reduction of Escherichia coli bacteria was obtained for the flow rate of 1 mL/hr, and no regrowth of bacteria was observed. Images from scanning electron microscope confirmed the formation of electroporation-induced nano-pore within the cell membrane. Through numerical simulation, it has been shown that sufficiently large electric field strength (3 kV/cm), required for bacterial electroporation, were generated using PDMS micropillars for an applied voltage of 300 V. Further, in this method of inactivation, there is no involvement of chemicals and the formation of harmful by-products is also minimum.

Keywords: electroporation, high-efficiency, inactivation, microfluidics, micropillar

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726 Using Hybrid Method for Inactivation of Microorganism and Enzymes in a Berry Juice

Authors: Golnoosh Torabian, P. Valtchev, F. Dehghani

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The need for efficient nutraceutical products has been dramatically changing the approach of the industrial processes. The development of novel mild processes is highly demanded for the production of such products; especially when both quality and safety need to be guaranteed during their long shelf life. Within this research, for the first time, we investigated the effect of supercritical carbon dioxide treatment for the inactivation of microbes and enzymes in a berry juice possessing therapeutic effect. We demonstrated that a complete inactivation of microbes can be achieved at optimized conditions of treatment. However, the bottle neck of the process was represented by the unpromising inactivation of the degradative enzyme by supercritical carbon dioxide treatment. However, complete enzyme inactivation was achieved by applying two strategies: the first was optimizing juicing method by adding a mechanical step and the second strategy was addition of natural inhibitors to the juice. Overall these results demonstrate that our hybrid process has a significant effect on the inactivation of microorganism and enzymes in the fresh juice. The developed process opens the possibility for the evolution of new products with optimal nutritional and sensorial characteristics, as well as offering a competitive cost and an environmentally friendly alternative for pasteurization and extension of shelf life in a wide range of natural therapeutic products.

Keywords: hybrid method, berry juice, pasteurization, enzymes inactivation

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725 Validation of Escherichia coli O157:H7 Inactivation on Apple-Carrot Juice Treated with Manothermosonication by Kinetic Models

Authors: Ozan Kahraman, Hao Feng

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Several models such as Weibull, Modified Gompertz, Biphasic linear, and Log-logistic models have been proposed in order to describe non-linear inactivation kinetics and used to fit non-linear inactivation data of several microorganisms for inactivation by heat, high pressure processing or pulsed electric field. First-order kinetic parameters (D-values and z-values) have often been used in order to identify microbial inactivation by non-thermal processing methods such as ultrasound. Most ultrasonic inactivation studies employed first-order kinetic parameters (D-values and z-values) in order to describe the reduction on microbial survival count. This study was conducted to analyze the E. coli O157:H7 inactivation data by using five microbial survival models (First-order, Weibull, Modified Gompertz, Biphasic linear and Log-logistic). First-order, Weibull, Modified Gompertz, Biphasic linear and Log-logistic kinetic models were used for fitting inactivation curves of Escherichia coli O157:H7. The residual sum of squares and the total sum of squares criteria were used to evaluate the models. The statistical indices of the kinetic models were used to fit inactivation data for E. coli O157:H7 by MTS at three temperatures (40, 50, and 60 0C) and three pressures (100, 200, and 300 kPa). Based on the statistical indices and visual observations, the Weibull and Biphasic models were best fitting of the data for MTS treatment as shown by high R2 values. The non-linear kinetic models, including the Modified Gompertz, First-order, and Log-logistic models did not provide any better fit to data from MTS compared the Weibull and Biphasic models. It was observed that the data found in this study did not follow the first-order kinetics. It is possibly because of the cells which are sensitive to ultrasound treatment were inactivated first, resulting in a fast inactivation period, while those resistant to ultrasound were killed slowly. The Weibull and biphasic models were found as more flexible in order to determine the survival curves of E. coli O157:H7 treated by MTS on apple-carrot juice.

Keywords: Weibull, Biphasic, MTS, kinetic models, E.coli O157:H7

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724 Inactivation of Rhodotorula spp. 74 with Cold Atmospheric Plasma

Authors: Zoran Herceg, Višnja Stulić, Tomislava Vukušić, Anet Režek Jambrak

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High voltage electrical discharge is a new technology used for inactivation of pathogen microorganisms. Pathogen yeasts can cause diseases in humans if they are ingested. Nowadays new technologies have become the focus of researching all over the world. Rhodotorula is known as yeast that can cause diseases in humans. The aim of this study was to examine whether the high voltage electrical discharge treatment generated in gas phase has an influence on yeast reduction and recovery of Rhodotorula spp 74 in pure culture. Rhodotorula spp. 74 was treated in 200 mL of model solution. Treatment time (5 and 10 min), frequency (60 and 90 Hz) and injected gas (air or argon 99,99%) were changed. Titanium high voltage needle was used as high voltage electrode (positive polarity) through which air or argon was injected at the gas flow of 0.6 L/min. Experimental design and statistical analyses were obtained by Statgraphics Centurion software (StatPoint Technologies, Inc., VA, USA). The best inactivation rate 1.7 log10 reduction was observed after the 10 min of treatment, frequency of 90 Hz and injected air. Also with a longer treatment time inactivation rate was higher. After the 24 h recovery of treated samples was observed. Therefore the further optimization of method is needed to understand the mechanism of yeasts inactivation and cells recovery after the treatment. Acknowledgements: The authors would like to acknowledge the support by Croatian Science Foundation and research project ‘Application of electrical discharge plasma for preservation of liquid foods’.

Keywords: rhodotorula spp. 74, electrical discharge plasma, inactivation, stress response

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723 Diffraction-Based Immunosensor for Dengue NS1 Virus

Authors: Harriet Jane R. Caleja, Joel I. Ballesteros, Florian R. Del Mundo

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The dengue fever belongs to the world’s major cause of death, especially in the tropical areas. In the Philippines, the number of dengue cases during the first half of 2015 amounted to more than 50,000. In 2012, the total number of cases of dengue infection reached 132,046 of which 701 patients died. Dengue Nonstructural 1 virus (Dengue NS1 virus) is a recently discovered biomarker for the early detection of dengue virus. It is present in the serum of the dengue virus infected patients even during the earliest stages prior to the formation of dengue virus antibodies. A biosensor for the dengue detection using NS1 virus was developed for faster and accurate diagnostic tool. Biotinylated anti-dengue virus NS1 was used as the receptor for dengue virus NS1. Using the Diffractive Optics Technology (dotTM) technique, real time binding of the NS1 virus to the biotinylated anti-NS1 antibody is observed. The dot®-Avidin sensor recognizes the biotinylated anti-NS1 and this served as the capture molecule to the analyte, NS1 virus. The increase in the signal of the diffractive intensity signifies the binding of the capture and the analyte. The LOD was found to be 3.87 ng/mL while the LOQ is 12.9 ng/mL. The developed biosensor was also found to be specific for the NS1 virus.

Keywords: avidin-biotin, diffractive optics technology, immunosensor, NS1

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722 Use of Thermosonication to Obtain Minimally Processed Mosambi Juice

Authors: Ruby Siwach, Manish Kumar, Raman Seth

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Extent of inactivation of pectin methylesterase (PME) in mosambi juice during thermal and thermosonication treatments was studied to obtain a minimally processed product. Effect of both treatments on cloud value, pH, titratable acidity, oBrix, and sensory attributes (flavour and taste) was studied. Thermal treatments (HT) were carried out at three temperatures 60, 70, and 80°C in a serological water bath for 5, 10, 15, and 20 min at each temperature. Thermosonication treatments (TS) were also given for same time-temperature combinations in water bath of a thermosonicator. Treated samples were stored in a deep freezer at 18°C for PME assay. PME activity of untreated sample was also assayed and residual PME activity and % loss in PME activity was calculated at each time-temperature combination. The extent of inactivation of PME increased with increase in treatment temperature and duration. Thermosonication treatments were found far more effective than thermal treatments of same time temperature combination in PME inactivation and retention of sensory attributes.

Keywords: pectin methylesterase, heat inactivation kinetics, thermosonication, thermal treatment

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721 Biosecurity Control Systems in Two Phases for Poultry Farms

Authors: M. Peña Aguilar Juan, E. Nava Galván Claudia, Pastrana Palma Alberto

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In this work was developed and implemented a thermal fogging disinfection system to counteract pathogens from poultry feces in agribusiness farms, to reduce mortality rates and increase biosafety in them. The control system consists of two phases for the conditioning of the farm during the sanitary break. In the first phase, viral and bacterial inactivation was performed by treating the stool dry cleaning, along with the development of a specialized product that foster the generation of temperatures above 55 °C in less than 24 hr, for virus inactivation. In the second phase, a process for disinfection by fogging was implemented, along with the development of a specialized disinfectant that guarantee no risk for the operators’ health or birds. As a result of this process, it was possible to minimize the level of mortality of chickens on farms from 12% to 5.49%, representing a reduction of 6.51% in the death rate, through the formula applied to the treatment of poultry litter based on oxidising agents used as antiseptics, hydrogen peroxide solutions, glacial acetic acid and EDTA in order to act on bacteria, viruses, micro bacteria and spores.

Keywords: innovation, triple helix, poultry farms, biosecurity

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720 A Comparative Study of Virus Detection Techniques

Authors: Sulaiman Al amro, Ali Alkhalifah

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The growing number of computer viruses and the detection of zero day malware have been the concern for security researchers for a large period of time. Existing antivirus products (AVs) rely on detecting virus signatures which do not provide a full solution to the problems associated with these viruses. The use of logic formulae to model the behaviour of viruses is one of the most encouraging recent developments in virus research, which provides alternatives to classic virus detection methods. In this paper, we proposed a comparative study about different virus detection techniques. This paper provides the advantages and drawbacks of different detection techniques. Different techniques will be used in this paper to provide a discussion about what technique is more effective to detect computer viruses.

Keywords: computer viruses, virus detection, signature-based, behaviour-based, heuristic-based

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719 Determination of Inactivation and Recovery of Saccharomyces cerevisiae Cells after the Gas-Phase Plasma Treatment

Authors: Z. Herceg, V. Stulic, T. Vukusic, A. Rezek Jambrak

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Gas phase plasma treatment is a new nonthermal technology used for food and water decontamination. In this study, we have investigated influence of the gas phase plasma treatment on yeast cells of S. cerevisiae. Sample was composed of 10 mL of yeast suspension and 190 mL of 0.01 M NaNO₃ with a medium conductivity of 100 µS/cm. Samples were treated in a glass reactor with a point- to-plate electrode configuration (high voltage electrode-titanium wire in the gas phase and grounded electrode in the liquid phase). Air or argon were injected into the headspace of the reactor at the gas flow of 5 L/min. Frequency of 60, 90 and 120 Hz, time of 5 and 10 min and positive polarity were defined parameters. Inactivation was higher with the applied higher frequency, longer treatment time and injected argon. Inactivation was not complete which resulted in complete recovery. Cellular leakage (260 nm and 280 nm) was higher with a longer treatment time and higher frequency. Leakage at 280 nm which defines a leakage of proteins was higher than leakage at 260 nm which defines a leakage of nucleic acids. The authors would like to acknowledge the support by Croatian Science Foundation and research project 'Application of electrical discharge plasma for preservation of liquid foods'.

Keywords: Saccharomyces cerevisiae, inactivation, gas-phase plasma treatment, cellular leakage

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718 Effect of Hydrogen Peroxide Concentration Produced by Cold Atmospheric Plasma on Inactivation of Escherichia Coli in Water

Authors: Zohreh Rashmei

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Introduction: Plasma inactivation is one of the emerging technologies in biomedical field and has been applied to the inactivation of microorganisms in water. The inactivation effect has been attributed to the presence of active plasma species, i.e. OH, O, O3, H2O2, UV and electric fields, generated by the discharge of plasma. Material and Method: To evaluate germicidal effects of plasma, the electric spark discharge device was used. After the effect of the plasma samples were collected for culture medium agar plate count. In addition to biological experiments, the concentration of hydrogen peroxide was also measured. Results: The results showed that Plasma is able to inactivate a high concentration of E. coli. After a short period of plasma radiation on the surface of water, the amount log8 reduced the microbial load. Starting plasma radiation on the surface of the water, the measurements show of production and increasing the amount of hydrogen peroxide in water. So that at the end of the experiment, the concentration of hydrogen peroxide to about 100 mg / l increased. Conclusion: Increasing the concentration of hydrogen peroxide is directly related to the reduction of microbial load. The results of E. coli culture in media containing certain concentrations of H2O2 showed that E. coli can not to grow in a medium containing more than 2/5 mg/l of H2O2. Surely we can say that the main cause of killing bacteria is a molecule of H2O2.

Keywords: plasma, hydrogen peroxide, disinfection, E. coli

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717 Zika Virus NS5 Protein Potential Inhibitors: An Enhanced in silico Approach in Drug Discovery

Authors: Pritika Ramharack, Mahmoud E. S. Soliman

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The re-emerging Zika virus is an arthropod-borne virus that has been described to have explosive potential as a worldwide pandemic. The initial transmission of the virus was through a mosquito vector, however, evolving modes of transmission has allowed the spread of the disease over continents. The virus already been linked to irreversible chronic central nervous system (CNS) conditions. The concerns of the scientific and clinical community are the consequences of Zika viral mutations, thus suggesting the urgent need for viral inhibitors. There have been large strides in vaccine development against the virus but there are still no FDA-approved drugs available. Rapid rational drug design and discovery research is fundamental in the production of potent inhibitors against the virus that will not just mask the virus, but destroy it completely. In silico drug design allows for this prompt screening of potential leads, thus decreasing the consumption of precious time and resources. This study demonstrates an optimized and proven screening technique in the discovery of two potential small molecule inhibitors of Zika virus Methyltransferase and RNA-dependent RNA polymerase. This in silico “per-residue energy decomposition pharmacophore” virtual screening approach will be critical in aiding scientists in the discovery of not only effective inhibitors of Zika viral targets, but also a wide range of anti-viral agents.

Keywords: NS5 protein inhibitors, per-residue decomposition, pharmacophore model, virtual screening, Zika virus

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716 Biopsy Proven Polyoma (BK) Virus in Saudi Kidney Recipients – Prevalence, Clinicopathological Features and Clinico-Pathological Correlations

Authors: Sarah Hamdan Al-Jahdali, Khaled Alsaad, Abdullah Al-Sayyari

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Objectives: To study the prevalence, clinicopathological features, risk factors and outcome of biopsy proven polyoma (BK) virus infection among Saudi kidney transplant recipients and compare them to negative BK virus group. Methods: We retrospectively reviewed the charts of all the patients with biopsy-proven polyoma (BK) virus infection in King Abdulaziz Medical City in Riyadh between 2005 and 2011. The details of clinical presentation, the indication for kidney biopsy, the laboratory findings at presentation, the natural history of the disease, thepathological findings, the prognosis as well as the response to therapy were all recorded. Results: Kidney biopsy was performed in 37 cases of unexplained graft dysfunction. BK virus was found in 10 (27%). Out of those 10, 3 (30%) ended with graft failure. BK virus occurred in all patients who received ATG induction therapy 100% versus 59.3% in the non BK virus patients (p=0.06). Furthermore, the risk of BK virus was much less in those who received acyclovir as an anti-viral prophylaxis as compared to those who did not receive it (p=0.01). Also, patients with BK virus weighed much less (mean 46.7±20.6 Kgs) than those without BK virus at time of transplantation (mean 64.3±12.1). Graft survival was better among deceased donor kidneys compared to living ones (P=0.016) and with older age (P=0.005). Conclusion: Our findings suggest the involvement of ATG induction therapy, the lack of antiviral prophylaxis therapy and lower weight at transplant as significant risk factors for the development of BK virus infection.

Keywords: BKVAN, BKV, kidney transpant, Saudi Arabia

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715 Survey of Potato Viral Infection Using Das-Elisa Method in Georgia

Authors: Maia Kukhaleishvili, Ekaterine Bulauri, Iveta Megrelishvili, Tamar Shamatava, Tamar Chipashvili

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Plant viruses can cause loss of yield and quality in a lot of important crops. Symptoms of pathogens are variable depending on the cultivars and virus strain. Selection of resistant potato varieties would reduce the risk of virus transmission and significant economic impact. Other way to avoid reduced harvest yields is regular potato seed production sampling and testing for viral infection. The aim of this study was to determine the occurrence and distribution of viral diseases according potato cultivars for further selection of virus-free material in Georgia. During the summer 2015- 2016, 5 potato cultivars (Sante, Laura, Jelly, Red Sonia, Anushka) at 5 different farms located in Akhalkalaki were tested for 6 different potato viruses: Potato virus A (PVA), Potato virus M (PVM), Potato virus S (PVS), Potato virus X (PVX), Potato virus Y (PVY) and potato leaf roll virus (PLRV). A serological method, Double Antibody Sandwich-Enzyme linked Immunosorbent Assay (DASELISA) was used at the laboratory to analyze the results. The result showed that PVY (21.4%) and PLRV (19.7%) virus presence in collected samples was relatively high compared to others. Researched potato cultivars except Jelly and Laura were infected by PVY with different concentrations. PLRV was found only in three potato cultivars (Sante, Jelly, Red Sonia) and PVM virus (3.12%) was characterized with low prevalence. PVX, PVA and PVS virus infection was not reported. It would be noted that 7.9% of samples were containing PVY/PLRV mix infection. Based on the results it can be concluded that PVY and PLRV infections are dominant in all research cultivars. Therefore significant yield losses are expected. Systematic, long-term control of potato viral infection, especially seed-potatoes, must be regarded as the most important factor to increase seed productivity.

Keywords: virus, potato, infection, diseases

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714 Eradication of Apple mosaic virus from Corylus avellana L. via Cryotherapy and Confirmation of Virus-Free Plants via Reverse Transcriptase Polymerase Chain Reaction

Authors: Ergun Kaya

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Apple mosaic virus (ApMV) is an ilarvirus causing harmful damages and product loses in many plant species. Because of xylem and phloem vessels absence, plant meristem tissues used for meristem cultures are virus-free, but sometimes only meristem cultures are not sufficient for virus elimination. Cryotherapy, a new method based on cryogenic techniques, is used for virus elimination. In this technique, 0.1-0.3mm meristems are excised from organized shoot apex of a selected in vitro donor plant and these meristems are frozen in liquid nitrogen (-196 °C) using suitable cryogenic technique. The aim of this work was to develop an efficient procedure for ApMV-free hazelnut via cryotherapy technique and confirmation of virus-free plants using Reverse Transcriptase-PCR technique. 100% virus free plantlets were obtained using droplet-vitrification method involved cold hardening in vitro cultures of hazelnut, 24 hours sucrose preculture of meristems on MS medium supplemented with 0.4M sucrose, and a 90 min PVS2 treatment in droplets.

Keywords: droplet vitrification, hazelnut, liquid nitrogen, PVS2

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713 Neutralizing Antibody Response against Inactivated FMDV Type O/IRN/2010 Vaccine by Electron Beam in BALB/C Mice

Authors: F. Motamedi Sedeh, Sh. Chahardoli, H. Mahravani, N. Harzandi, M. Sotoodeh, S. K. Shafaei

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Foot-and-mouth disease virus (FMDV) is the most economically important disease of livestock. The aim of the study is inactivation of FMD virus type O/IRN/2010 by electron beam without antigenic changes as electron radio vaccine. The BALB/C mice were divided into three groups, each group containing five mice. Three groups of mice were inoculated with conventional vaccine and electron beam irradiated vaccine FMDV type O/IRN/2010 subcutaneously three weeks interval, the final group as negative control. The sera were separated from the blood samples of mice 14 days after last vaccination and tested for the presence of antibodies against FMDV type O/IRN/2010 by serum neutralization test. The Serum Neutralization Test (SNT) was carried out and antibody titration was calculated according to the Kraber protocol. The results of the SNT in three groups of mice showed the titration of neutralizing antibody in the vaccinated mice groups; electron radio vaccine and conventional vaccine were significantly higher than negative control group (P<0.05). Therefore, the radio vaccine is a good candidate to immunize animals against FMDV type O/IRN/2010.

Keywords: FMDV type O/IRN/2010, neutralizing antibody response, electron beam, radio vaccine

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712 Global Analysis of HIV Virus Models with Cell-to-Cell

Authors: Hossein Pourbashash

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Recent experimental studies have shown that HIV can be transmitted directly from cell to cell when structures called virological synapses form during interactions between T cells. In this article, we describe a new within-host model of HIV infection that incorporates two mechanisms: infection by free virions and the direct cell-to-cell transmission. We conduct the local and global stability analysis of the model. We show that if the basic reproduction number R0 1, the virus is cleared and the disease dies out; if R0 > 1, the virus persists in the host. We also prove that the unique positive equilibrium attracts all positive solutions under additional assumptions on the parameters.

Keywords: HIV virus model, cell-to-cell transmission, global stability, Lyapunov function, second compound matrices

Procedia PDF Downloads 477
711 Impact of Totiviridae L-A dsRNA Virus on Saccharomyces Cerevisiae Host: Transcriptomic and Proteomic Approach

Authors: Juliana Lukša, Bazilė Ravoitytė, Elena Servienė, Saulius Serva

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Totiviridae L-A virus is a persistent Saccharomyces cerevisiae dsRNA virus. It encodes the major structural capsid protein Gag and Gag-Pol fusion protein, responsible for virus replication and encapsulation. These features also enable the copying of satellite dsRNAs (called M dsRNAs) encoding a secreted toxin and immunity to it (known as killer toxin). Viral capsid pore presumably functions in nucleotide uptake and viral mRNA release. During cell division, sporogenesis, and cell fusion, the virions remain intracellular and are transferred to daughter cells. By employing high throughput RNA sequencing data analysis, we describe the influence of solely L-A virus on the expression of genes in three different S. cerevisiae hosts. We provide a new perception into Totiviridae L-A virus-related transcriptional regulation, encompassing multiple bioinformatics analyses. Transcriptional responses to L-A infection were similar to those induced upon stress or availability of nutrients. It also delves into the connection between the cell metabolism and L-A virus-conferred demands to the host transcriptome by uncovering host proteins that may be associated with intact virions. To better understand the virus-host interaction, we applied differential proteomic analysis of virus particle-enriched fractions of yeast strains that harboreither complete killer system (L-A-lus and M-2 virus), M-2 depleted orvirus-free. Our analysis resulted in the identification of host proteins, associated with structural proteins of the virus (Gag and Gag-Pol). This research was funded by the European Social Fund under the No.09.3.3-LMT-K-712-19-0157“Development of Competences of Scientists, other Researchers, and Students through Practical Research Activities” measure.

Keywords: totiviridae, killer virus, proteomics, transcriptomics

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710 Chikungunya Virus Infection among Patients with Febrile Illness Attending University of Maiduguri Teaching Hospital, Nigeria

Authors: Abdul-Dahiru El-Yuguda, Saka Saheed Baba, Tawa Monilade Adisa, Mustapha Bala Abubakar

Abstract:

Background: Chikungunya (CHIK) virus, a previously anecdotally described arbovirus, is now assuming a worldwide public health burden. The CHIK virus infection is characterized by potentially life threatening and debilitating arthritis in addition to the high fever, arthralgia, myalgia, headache and rash. Method: Three hundred and seventy (370) serum samples were collected from outpatients with febrile illness attending University of Maiduguri Teaching Hospital, Nigeria, and was used to detect for Chikungunya (CHIK) virus IgG and IgM antibodies using the Enzyme Linked Immunosorbent Assays (ELISAs). Result: Out of the 370 sera tested, 39 (10.5%) were positive for presence of CHIK virus antibodies. A total of 24 (6.5%) tested positive for CHIK virus IgM only while none (0.0%) was positive for presence of CHIK virus IgG only and 15 (4.1%) of the serum samples were positive for both IgG and IgM antibodies. A significant difference (p<0.0001) was observed in the distribution of CHIK virus antibodies in relation to gender. The males had prevalence of 8.5% IgM antibodies as against 4.6% observed in females. On the other hand 4.6% of the females were positive for concurrent CHIK virus IgG and IgM antibodies when compared to a prevalence of 3.4% observed in males. Only the age groups ≤ 60 years and the undisclosed age group were positive for presence of CHIK virus IgG and/or IgM antibodies. No significant difference (p>0.05) was observed in the seasonal prevalence of CHIK virus antibodies among the study subjects Analysis of the prevalence of CHIK virus antibodies in relation to clinical presentation (as observed by Clinicians) of the patients revealed that headache and fever were the most frequently encountered ailments. Conclusion: The CHIK virus IgM and concurrent IgM and IgG antibody prevalence rates of 6.5% and 4.1% observed in this study indicates a current infection and the lack of IgG antibody alone observed shows that the infection is not endemic but sporadic. Recommendation: Further studies should be carried to establish the seasonal prevalence of CHIK virus infection vis-à-vis vector dynamics in the study area. A comprehensive study need to be carried out on the molecular characterization of the CHIK virus circulating in Nigeria with a view to developing CHIK virus vaccine.

Keywords: Chikungunya virus, IgM and IgG antibodies, febrile patients, enzyme linked immunosorbent assay

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709 Inactivation of Root-Knot Nematode Eggs Meloidogyne enterolobii in Irrigation Water Treated with Ozone

Authors: I. A. Landa-Fernandez, I. Monje-Ramirez, M. T. Orta-Ledesma

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Every year plant-parasitic nematodes diminish the yield of high-value crops worldwide causing important economic losses. Currently, Meloidogyne enterolobii has increased its importance due to its high aggressiveness, increasing geographical distribution and host range. Root-knot nematodes inhabit the rhizosphere soil around plant roots. However, they can come into contact with irrigation water. Thus, plant-parasitic nematodes can be transported by water, as eggs or juveniles. Due to their high resistance, common water disinfection methods are not effective for inactivating these parasites. Ozone is the most effective disinfectant for microbial inactivation. The objective of this study is to demonstrate that ozone treatment is an alternative method control in irrigation water of the root-knot nematode M. enterolobii. It has been shown that ozonation is an effective treatment for the inactivation of protozoan cysts and oocysts (Giardia and Cryptosporidium) and for other species of the genus Meloidogyne (M. incognita), but not for the enterolobii specie. In this study, the strain of M. enterolobii was isolated from tomatoes roots. For the tests, eggs were used and were inoculated in water with similar characteristics of irrigation water. Subsequently, the disinfection process was carried out in an ozonation unit. The performance of the treatments was evaluated through the egg's viability by assessing its structure by optical microscopy. As a result of exposure to ozone, the viability of the nematode eggs was reduced practically in its entirety; with dissolved ozone levels in water close to the standard concentration (equal to 0.4 mgO₃/L), but with high contact times (greater than 4 min): 0.2 mgO₃/L for 15 minutes or 0.55 mgO₃/L for 10 minutes. Additionally, the effect of temperature, alkalinity and organic matter of the water was evaluated. Ozonation is effective and a promising alternative for the inactivation of nematodes in irrigation water, which could contribute to diminish the agricultural losses caused by these organisms.

Keywords: inactivation process, irrigation water treatment, ozonation, plant-parasite nematodes

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708 Inactivation and Stress Response of Salmonella enterica Serotype Typhimurium lt21 upon Cold Gas-Phase Plasma Treatment

Authors: Zoran Herceg, Tomislava Vukušić, Anet Režek Jambrak, Višnja Stulić

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Today one of the greatest challenges are directed to the safety of food supply. If food pathogens are ingested they can cause human illnesses. Because of that new technologies that are effective in microbial reduction are developing to be used in food industries. One of such technology is cold gas phase plasma. Salmonella enterica was studied as one of the pathogenes that can be found in food. The aim of this work was to examine the inactivation rate and stress response of plasma treated cells of Salmonella enterica inoculated in apple juice. After the treatment cellular leakage, phenotypic changes in plasma treated cells-biofilm formation and degree of recovery were conducted. Sample volume was inoculated with 5 mL of pure culture of Salmonella enterica and 15 mL of apple juice. Statgraphics Centurion software (StatPoint Technologies, Inc., VA, USA) was used for experimental design and statistical analyses. Treatment time (1, 3, 5 min) and gas flow (40, 60, 80 L/min) were changed. Complete inactivation and 0 % of recovery after the 48 h was observed at these experimental treatments: 3 min; 40 L/min, 3 min; 80 L/min, 5 min; 40 L/min. Biofilm reduction was observed at all treated samples. Also, there was an increase in cellular leakage with a longer plasma treatment. Although there were a significant reduction and 0 % of recovery after the plasma treatments further investigation of the method is needed to clarify whether there are sensorial, physical and chemical changes in juices after the plasma treatment. Acknowledgments: The authors would like to acknowledge the support by Croatian Science Foundation and research project 'Application of electrical discharge plasma for the preservation of liquid foods'.

Keywords: salmonella enterica serotype typhimurium lt21, gas-phase plasma treatment, inactivation, stress response

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707 Growth of Metal Oxide (Tio2/Ag) Thin Films Sputtered by Hipims Effective in Bacterial Inactivation: Plasma Chemistry and Energetic

Authors: O. Baghriche, A. Zertal, C. Pulgarin, J. Kiwi, R. Sanjines

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High-Power Impulse Magnetron Sputtering (HIPIMS) is a technology that belongs to the field of Ionized PVD of thin films. This study shows the first complete report on ultrathin TiO2/Ag nano-particulate films sputtered by highly ionized pulsed plasma magnetron sputtering (HIPIMS) leading to fast bacterial loss of viability. The Ag and the TiO2/Ag sputtered films induced complete Escherichia coli inactivation in the dark, which was not observed in the case of TiO2. When Ag was present, the bacterial inactivation was accelerated under low intensity solar simulated light and this has implications for a potential for a practical technology. The design, preparation, testing and surface characterization of these innovative films are described in this study. The HIPIMS sputtered composite films present an appreciable savings in metals compared to films obtained by conventional sputtering methods. HIPIMS sputtering induces a strong interaction with the rugous polyester 3-D structure due to the higher fraction of the Ag-ions (M+) attained in the magnetron chamber. The immiscibility of Ag and TiO2 in the TiO2/Ag films is shown by High Angular Dark Field (HAADF) microscopy. The ionization degree of the film forming species is significantly increased and film growth is assisted by an intense ion flux. Reports have revealed the significant enhancement of the film properties as the HIPIMS technology is used. However, a decrease of the deposition rate, as compared to the conventional DC magnetron sputtering Pulsed (DCMSP) process is commonly observed during HIPIMS.

Keywords: E. coli, HIPIMS, inactivation bacterial, sputtering

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706 Physicochemical Properties and Thermal Inactivation of Polyphenol Oxidase of African Bush Mango (Irvingia Gabonensis) Fruit

Authors: Catherine Joke Adeseko

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Enzymatic browning is an economically important disorder that degrades organoleptic properties and prevent the consumer from purchasing fresh fruit and vegetables. Prevention and control of enzymatic browning in fruit and its product is imperative. Therefore, this study sought to investigate the catalytic effect of polyphenol oxidase (PPO) in the adverse browning of African bush mango (Irvingia gabonensis) fruit peel and pulp. PPO was isolated and purified, and its physicochemical properties, such as the effect of pH with SDS, temperature, and thermodynamic studies, which invariably led to thermal inactivation of purified PPO at 80 °C, were evaluated. The pH and temperature optima of PPO were found at 7.0 and 50, respectively. There was a gradual increase in the activity of PPO as the pH increases. However, the enzyme exhibited a higher activity at neutral pH 7.0, while enzymatic inhibition was observed at acidic region, pH 2.0. The presence of SDS at pH 5.0 downward was found to inhibit the activity of PPO from the peel and pulp of I. gabonensis. The average value of enthalpy (ΔH), entropy (ΔS), and Gibbs free energy (ΔG) obtained at 20 min of incubation and temperature 30 – 80 °C were respectively 39.93 kJ.mol-1, 431.57 J.mol-1 .K-1 and -107.99 kJ.mol-1 for peel PPO, and 37.92 kJ.mol-1, -442.51J.mol-1.K-1, and -107.22 kJ.mol-1 for pulp PPO. Thermal inactivation of PPO from I. gabonensis exhibited a reduction in catalytic activity as the temperature and duration of heat inactivation increases using catechol, reflected by an increment in k value. The half-life of PPO (t1/2) decreases as the incubation temperature increases due to the instability of the enzyme at high temperatures and was higher in pulp than peel. Both D and Z values decrease with increase in temperature. The information from this study suggests processing parameters for controlling PPO in the potential industrial application of I. gabonensis fruit in order to prolong the shelf-life of this fruit for maximum utilization.

Keywords: enzymatic, browning, characterization, activity

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705 DNA Vaccine Study against Vaccinia Virus Using In vivo Electroporation

Authors: Jai Myung Yang, Na Young Kim, Sung Ho Shin

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The adverse reactions of current live smallpox vaccines and potential use of smallpox as a bioterror weapon have heightened the development of new effective vaccine for this infectious disease. In the present study, DNA vaccine vector was produced which was optimized for expression of the vaccinia virus L1 antigen in the mouse model. A plasmid IgM-tL1R, which contains codon-optimized L1R gene, was constructed and fused with an IgM signal sequence under the regulation of a SV40 enhancer. The expression and secretion of recombinant L1 protein was confirmed in vitro 293 T cell. Mice were administered the DNA vaccine by electroporation and challenged with vaccinia virus. We observed that immunization with IgM-tL1R induced potent neutralizing antibody responses and provided complete protection against lethal vaccinia virus challenge. Isotyping studies reveal that immunoglobulin G2 (IgG2) antibody predominated after the immunization, indicative of a T helper type 1 response. Our results suggest that an optimized DNA vaccine, IgM-tL1R, can be effective in stimulating anti-vaccinia virus immune response and provide protection against lethal orthopoxvirus challenge.

Keywords: DNA vaccine, electroporation, L1R, vaccinia virus

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704 Effects of Ultraviolet Treatment on Microbiological Load and Phenolic Content of Vegetable Juice

Authors: Kubra Dogan, Fatih Tornuk

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Due to increasing consumer demand for the high-quality food products and awareness regarding the health benefits of different nutrients in food minimal processing becomes more popular in modern food preservation. To date, heat treatment is often used for inactivation of spoilage microorganisms in foods. However, it may cause significant changes in the quality and nutritional properties of food. In order to overcome the detrimental effects of heat treatment, several alternatives of non-thermal microbial inactivation processes have been investigated. Ultraviolet (UV) inactivation is a promising and feasible method for better quality and longer shelf life as an alternative to heat treatment, which aims to inhibit spoilage and pathogenic microorganisms and to inactivate the enzymes in vegetable juice production. UV-C is a sub-class of UV treatment which shows the highest microcidal effect between 250-270 nm. The wavelength of 254 nm is used for the surface disinfection of certain liquid food products such as vegetable juice. Effects of UV-C treatment on microbiological load and quality parameter of vegetable juice which is a mix of celery, carrot, lemon and orange was investigated. Our results showed that storing of UV-C applied vegetable juice for three months, reduced the count of TMAB by 3.5 log cfu/g and yeast-mold by 2 log cfu/g compared to control sample. Total phenolic content was found to be 514.3 ± 0.6 mg gallic acid equivalent/L, and there wasn’t a significant difference compared to control. The present work suggests that UV-C treatment is an alternative method for disinfection of vegetable juice since it enables adequate microbial inactivation, longer shelf life and has minimal effect on degradation of quality parameters of vegetable juice.

Keywords: heat treatment, phenolic content, shelf life, ultraviolet (UV-C), vegetable juice

Procedia PDF Downloads 171