Search results for: Chlorella virus

Authors: Numfon Rakkhumkaew, Takeru Kawasaki, Makoto Fujie, Takashi Yamada

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Chlorella viruses or chloroviruses contain a gene that encodes a function for chitin synthesis, which is expressed early in viral infection to produce chitin polysaccharide, a polymer of β-1, 4-linked GlcNAc, on the outside of Chlorella cell wall. Interestingly, chlorovirus system is an eco-friendly system which converses CO2 and solar energy from the environment into useful materials. However, infected Chlorella cells are lysed at the final stage of viral infection, and this phenomenon is caused the breaking down of polysaccharide. To postpone the lysing period and prolong the synthesis of chitin polysaccharide on cells, the slow growing virus incorporated with aphidicolin treatment, an inhibitor of DNA synthesis, was investigated. In this study, a total of 25 virus isolates from water samples in Japan region were analyzed for CHS (the gene for CH synthase) gene by PCR (polymerase chain reaction). The accumulation and appearance of chitin polysaccharide on infected cells were detected by biotinylated chitin-binding proteins WGA (wheat germ agglutinin)-biotin for chitin in conjunction with avidin-Cy 2 or Cy 3 and investigated by fluorescence microscopy, observed as green or yellow fluorescence over the cell surface. Among all chlorovirus isolates, cells infected with CNF1 revealed the accumulation of chitin over the cell surface within 30 min p.i. and continued to accumulate on cells until 4 h p.i. before cell lyses which was 1.6 times longer accumulation period than cells infected with CVK2 (prototype virus). Furthermore, addition of aphidicolin could extend the chitin accumulation on cells infected with CNF1 until 8 h p.i. before cell lyses. Whereas, CVK2-infected cells treated with aphidicolin could prolong the chitin synthesis only for 6 h p.i. before cell lyses. Therefore, chitin synthesis by Chlorella-virus system could be prolonged by using slow-growing viral isolates and with aphidicolin.

Keywords: chitin, chlorovirus, Chlorella virus, aphidicolin

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Authors: Ratanaporn Leesing, Thidarat Papone, Mutiyaporn Puangbut

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Mixotrophic cultivation of the isolated freshwater microalgae Chlorella sp. KKU-S2 in batch shake flask for biomass and lipid productions, different concentration of glucose as carbon substrate, different nitrogen source and concentrations were investigated. Using 1.0g/L of NaNO3 as nitrogen source, the maximum biomass yield of 10.04g/L with biomass productivity of 1.673g/L d was obtained using 40g/L glucose, while a biomass of 7.09, 8.55 and 9.45g/L with biomass productivity of 1.182, 1.425 and 1.575g/L d were found at 20, 30 and 50g/L glucose, respectively. The maximum lipid yield of 3.99g/L with lipid productivity of 0.665g/L d was obtained when 40g/L glucose was used. Lipid yield of 1.50, 3.34 and 3.66g/L with lipid productivity of 0.250, 0.557 and 0.610g/L d were found when using the initial concentration of glucose at 20, 30 and 50g/L, respectively. Process product yield (YP/S) of 0.078, 0.119, 0.158 and 0.094 were observed when glucose concentration was 20, 30, 40 and 50 g/L, respectively. The results obtained from the study shows that mixotrophic culture of Chlorella sp. KKU-S2 is a desirable cultivation process for microbial lipid and biomass production.

Keywords: mixotrophic cultivation, microalgal lipid, Chlorella sp. KKU-S2

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Authors: Abdullah M. Alzahrani

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The unicellular alga Chlorella vulgaris was isolated from Al-Asfar Lake, which is located in the Al-Ahsa province of Saudi Arabia. Two different isolates were sub-cultured under laboratory conditions. The wild type was grown under a regular concentration of copper, whereas the other isolate was grown under a progressively increasing copper concentration. An Inter Simple Sequence Repeats (ISSR) analysis was performed using DNA isolated from the wild type and tolerant strains. The sum of the scored bands of the wild type was 155, with 100 (64.5%) considered to be polymorphic bands, whereas the resistant strain displayed 147 bands, with 92 (62.6%) considered to be polymorphic bands. The sum of the scored bands of a mixed sample was 117 bands, of which only 4 (3.4%) were considered to be polymorphic. The average Nei's genetic diversity (h) and Shannon-Weiner diversity indices (I) were 0.3891 and 0.5394, respectively. These results clearly indicate that the adaptation to a high level of copper in Chlorella vulgaris is not merely physiological but rather driven by modifications at the genomic level.

Keywords: chlorella vulgaris, copper tolerance, genetic diversity, green algae

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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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Authors: M. Alsull

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In this study, Tetraselmis sp. and Chlorella sp. isolated from Penang National Park coastal waters, Malaysia, and cultivated under combined various laboratory conditions (temperature, light and nitrogen limitation and starvation). Growth rate, dry weight, chlorophyll a content, total lipids content and total carbohydrates content were estimated at mid-exponential growth phase. Tetraselmis sp. and Chlorella sp. showed remarkably decrease in growth rate, chlorophyll a content and dry weight when maintained under nitrogen limitation and starvation conditions, as well as when grown under 12:12 h light, dark regime conditions. Chlorella sp. showed ability to counter the fluctuation in temperature with no significant effects on the measured parameters; in contrast, Tetraselmis sp. showed a decrease in growth rate, chlorophyll a content and dry weight when grown under 15±1˚C temperature. Cultures maintained under nitrogen full concentration, and 24 h light regime showed decrease in total lipids content, compared with 12:12 h light, dark cycle regime, in the two tested species.

Keywords: microalgae, biochemical composition, temperature, light, nitrogen limitation

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Authors: Osman Kök, İlhami̇ Tüzün, Yaşar Aluç

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This study was conducted to explore the effect of direct current (DC) applications on the growth of microalgae Chlorella vulgaris KKU71, isolated from highly saline freshwater. Experiments were implemented based upon the cross-combinations of both the intensity and duration of electric applications, generating a full factorial design of 10V, 20V, 30V, and 5s, 30s, 60s, respectively. Growth parameters of cultures were monitored on Optical Density (OD), Cell Count (CC), Chlorophyll-a, b (Chl-a, b), and Total Carotenoids (TCar). All DC-assisted treatments stimulated the growth and thus led to higher values of growth parameters such as OD, CC, Chl-a, and TCar. Monotonically increasing with the intensity and duration of DC applications, wet and dry biomass yields of the harvested algae reached their highest level at 30V-60s in all sets of treatments. In addition, this increase between DC applications was listed as C(control)<10V<20V<30V and C<5s<30s<60s. As a result, direct current applications increased the biomass.

Keywords: Chlorella Vulgaris, direct current, growth, biomass

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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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Authors: Jyoti Singh, Swati Dubey, Mukta Singh, R. P. Singh

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The freshwater microalgae Chlorella sp. is alluring considerable interest as a source for biofuel production due to its fast growth rate and high lipid content. Under nitrogen limited conditions, they can accumulate significant amounts of lipids. Thus, it is important to gain insight into the molecular mechanism of their lipid metabolism. In this study under nitrogen limited conditions, regular pattern of growth characteristics lipid accumulation and gene expression analysis of key regulatory genes of lipid biosynthetic pathway were carried out in microalgae Chlorella sp 3. Our results indicated that under nitrogen limited conditions there is a significant increase in the lipid content and lipid productivity, achieving 44.21±2.64 % and 39.34±0.66 mg/l/d at the end of the cultivation, respectively. Time-course transcript patterns of lipid biosynthesis genes i.e. acetyl coA carboxylase (accD) and diacylglycerol acyltransferase (dgat) showed that during late log phase of microalgae Chlorella sp.3 both the genes were significantly up regulated as compared to early log phase. Moreover, the transcript level of the dgat gene is two-fold higher than the accD gene. The results suggested that both the genes responded sensitively to the nitrogen limited conditions during the late log stage, which proposed their close relevance to lipid biosynthesis. Further, this transcriptome data will be useful for engineering microalgae species by targeting these genes for genetic modification to improve microalgal biofuel quality and production.

Keywords: biofuel, gene, lipid, microalgae

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Authors: Pramod Kumar, A. V. N. Swamy, C. V. Sowjanya, C. V. Ramachandra Murthy

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Biosorption is one of the effective methods for the removal of heavy metal ions from aqueous solutions. Results are presented showing the sorption of Pb(II) from solutions by biomass of commonly available marine algae Chlorella sp. The ability of marine algae Chlorella pyrenoidosa to remove heavy metal ion (Pb(II)) from aqueous solutions has been studied in this work. The biosorption properties of the biosorbent like equilibrium agitation time, optimum pH, temperature and initial solute concentration were investigated on metal uptake by showing respective profiles. The maximum metal uptake was found to be 10.27 mg/g. To achieve this metal uptake, the optimum conditions were found to be 30 min as equilibrium agitation time, 4.6 as optimum pH, 60 ppm of initial solute concentration. Lead concentration is determined by atomic absorption spectrometer. The process was found to be well fitted for pseudo-second order kinetics.

Keywords: biosorption, heavy metal ions, agitation time, metal uptake, aqueous solution

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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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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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Authors: Adrian Garrido Sanchis, Richard Pashley

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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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Authors: Gian Carlo F. Maliwat, Stephanie F. Velasquez, Janice A. Ragaza

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A 50-day growth trial was conducted to evaluate the efficacy of Chlorella vulgaris (Beijerinck) as an ingredient in the diets of giant freshwater prawn Macrobrachium rosenbergii (De Man) postlarvae (PL30). Immune response (total haemocyte count and prophenoloxidase activity) was also assessed by subjecting postlarvae to a challenge test against Aeromonas hydrophila (Chester) for 14 days. Isonitrogenous and iso-lipidic test diets were prepared using a fishmeal-based-positive control diet (D0) and four basal diets with inclusion levels of 2% (D2), 4% (D4), 6% (D6) and 8% (D8) C. vulgaris. Postlarvae of M. rosenbergii were randomly stocked (mean initial body weight of 0.19 ± 0.02 g) in 30-L tanks in three replicates per dietary treatment for evaluation of growth performance. Another set of postlarvae (mean initial body weight of 1.25 ± 0.02 g) was randomly distributed in 95-L tanks in three replicates per dietary treatment for the assessment of immune response. Results showed that specific growth rate was significantly higher (P < 0.05) in postlarvae fed D4 and D6. Variations in values for carcass protein, lipid, moisture, and ash were also evident. Postlarvae fed diets with Chlorella showed increased prophenol oxidase activity and total haemocyte counts. Moreover, the survival rate after challenge with A. hydrophila was significantly increased (P < 0.05). Inclusion of C. vulgaris in diets enhanced immune response and resistance of M. rosenbergii postlarvae against A. hydrophila infection.

Keywords: Chlorella vulgaris, haemocyte count, Macrobrachium rosenbergii, prophenoloxidase activity

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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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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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Authors: Al-Saboonchi Azhar

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The effects of Iraqi crude oil on growth of three freshwater algae (Chlorella vulgaris Beij., Scenedesmus acuminatus (Lag.) Chodat. and Oscillatoria princeps Vauch.) were investigated, basing on it's biomass expressed as Chl.a. Growth rate and doubling time of the cell were calculated. Results showed that growth rate and species survival varied with concentrations of crude oil and species type. Chlorella vulgaris and Scenedesmus acuminatus were more sensitive in culture containing crude oil as compared with Oscillatoria princeps cultures. The growth of green algae were significantly inhibited in culture containing (5 mg/l) crude oil, while the growth of Oscillatoria princeps reduced in culture containing (10 mg/l) crude oil.

Keywords: algae, crude oil, green algae, Cyanobacteria

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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

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

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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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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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Authors: Dilek Zorlu Kaya, Sena Çenesiz, Utku Duran

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Infectious pancreatic necrosis virus is a disease of great concern in aquaculture, causing mortality of 80 - 90% of the stocks in salmonid production. We aimed to investigate the efficacy of quercetin on oxidant and antioxidant parameters of infectious pancreatic necrosis virus, which is important for fish farming and economy in vitro. Quercetin experimental model was used in the cell culture of Oncorhynchus mykiss infected with infectious pancreatic necrosis virus. Malondialdehyde, ceruloplasmin, total oxidant capacity, total antioxidant levels, and glutathione-peroxidase were measured in the samples. As a result of the study, it was observed that quercetin can minimize the damage caused by scavenging free radicals in cells infected with infectious pancreatic necrosis virus. Thus, we think that an important development can be achieved for fish farming and the economy.

Keywords: IPNV, oncorhynchus mykiss, TAS, TOS, quercetin

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Authors: Titin Handayani, Adi Mulyanto, Fajar Eko Priyanto

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It is generally recognized, that algae could be an interesting option for reducing CO₂ emissions. Based on light and CO₂, algae can be used for the production various economically interesting products. Current algae cultivation techniques, however, still present a number of limitations. Efficient feeding of CO₂, especially on a large scale, is one of them. Current methods for CO₂ feeding to algae cultures rely on the sparging pure CO₂ or directly from flue gas. The limiting factor in this system is the solubility of CO₂ in water, which demands a considerable amount of energy for an effective gas to liquid transfer and leads to losses to the atmosphere. Due to the current ineffective methods for CO₂ introduction into algae ponds very large surface areas would be required for enough ponds to capture a considerable amount of the CO₂. The purpose of this study is to assess technology to capture carbon dioxide (CO₂) emissions generated by industry by utilizing of microalgae Chlorella sp. The microalgae were cultivated in a bioreactor culture pond raceway type. The result is expected to be useful in mitigating the effects of greenhouse gases in reducing the CO₂ emissions. The research activities include: (1) Characterization of boiler flue gas, (2) Operation of culture pond, (3) Sampling and sample analysis. The results of this study showed that the initial assessment absorption of the flue gas by microalgae using 1000 L raceway pond completed by heat exchanger were quite promising. The transfer of CO₂ into the pond culture system was run well. This identified from the success of cooling the boiler flue gas from the temperature of about 200 °C to below ambient temperature. Except for the temperature, the gas bubbles into the culture media were quite fine. Therefore, the contact between the gas and the media was well performed. The efficiency of CO₂ absorption by Chlorella sp reached 6.68 % with an average CO₂ loading of 0.29 g/L/day.

Keywords: Chlorella sp., CO2 emission, heat exchange, microalgae, milk industry, raceway pond

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Authors: Osagiede Efosa Kelvin

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The Ebola virus disease (EVD); also Ebola hemorrhagic fever, is a disease of humans and other primates caused by Ebola viruses. Signs and symptoms typically start between two days and three weeks after contracting the virus as a fever, sore throat, muscle pain, and headaches. Then, vomiting, diarrhoea and rash usually follow, along with decreased function of the liver and kidneys. At this time, some people begin to bleed both internally and externally. The first death in Nigeria was reported on 25 July 2014: a Liberian-American with Ebola flew from Liberia to Nigeria and died in Lagos soon after arrival. As part of the effort to contain the disease, possible contacts were monitored –353 in Lagos and 451 in Port Harcourt On 22 September, the World Health Organisation reported a total of 20 cases, including eight deaths. The WHO's representative in Nigeria officially declared Nigeria Ebola-free on 20 October after no new active cases were reported in the follow-up contact. This paper looks at the Ebola Virus in general and the measures taken by Nigeria to combat its spread.

Keywords: Ebola virus, hemorrhagic fever, Nigeria, outbreak

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Authors: Anjana Goel, Ashok Kumar Bhatia

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Ocimum sanctum, a well known medicinal plant is used for various alignments in Ayurvedic medicines. It was found to be effective in treating the humans suffering from different viral infections like chicken pox, small pox, measles and influenza. In addition, curative effect of the plant in malignant patients was also reported. In the present study, leaves of this plant were screened against animal viruses i.e. Bovine Herpes Virus-type-1 (BHV-1), Foot and Mouth disease virus (FMDV) and Newcastle Disease Virus (NDV). BHV-1 and FMDV were screened in MDBK and BHK cell lines respectively using cytopathic inhibition test. While NDV was propagated in chick embryo fibroblast culture and tested by haemagglutination inhibition test. Maximum non toxic dose of aqueous extract of Ocimum sanctum leaves was calculated by MTT assay in all the cell cultures and nontoxic doses were used for antiviral activity against viruses. 98.4% and 85.3% protection were recorded against NDV and BHV-1 respectively. However, Ocimum sanctum extract failed to show any inhibitory effect on the cytopathic effect caused by FMD virus. It can be concluded that Ocimum sanctum is a very effective remedy for curing viral infections in animals also.

Keywords: bovine herpes virus-type-1, foot and mouth disease virus, newcastle disease virus, Ocimum sanctum

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Authors: Vallejo Michael, Acuña Edgar, Roa Juan David, Peñuela Rosa, Parra Alejandra, Casallas Daniela, Rodriguez Sheyla

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Congenital Zika Virus Syndrome is an entity composed by a variety of birth defects presented in newborns that have been exposed to the Zika Virus during pregnancy. The syndrome characteristic features are severe microcephaly, cerebral tissue abnormalities, ophthalmological abnormalities such as uveitis and chorioretinitis, arthrogryposis, clubfoot deformity and muscular tone abnormalities. The confirmatory test is the Reverse transcription polymerase chain reaction (RT-PCR) associated to the physical findings. Here we present the case of a newborn with microcephaly whose mother presented a confirmed Zika Virus infection during the third trimester of pregnancy, despite of the evident findings and the history of Zika infection the RT-PCR in amniotic and cerebrospinal fluid of the newborn was negative. RT-PCR has demonstrated a low sensibility in samples with low viral loads, reason why, we propose a clinical diagnosis in patients with clinical history of Zika Virus infection during pregnancy accompanied by evident clinical manifestations of the child.

Keywords: congenital, Zika virus, microcephaly, reverse transcriptase polymerase chain reaction

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Authors: Dmitry Nosik, Nickolay Nosik, Elli Kaplina, Olga Lobach, Marina Chataeva, Lev Rasnetsov

Abstract:

Background and aims: Human Immunodeficiency Virus (HIV) infection is very often associated with Herpes Simplex Virus (HSV) infection but HIV patients are treated with a cocktail of antiretroviral drugs which are toxic. The use of an antiviral drug which will be active against both viruses like ferrovir found in our previous studies is rather actual. Earlier we had shown that Fullerene poly-amino capronic acid (FPACA) was active in case of monoinfection of HIV-1 or HSV-1. The aim of the study was to analyze the efficiency of FPACA against mixed infection of HIV and HSV. Methods: The peripheral blood lymphocytes, CEM, MT-4 cells were simultaneously infected with HIV-1 and HSV-1. FPACA was added 1 hour before infection. Cells viability was detected by MTT assay, virus antigens detected by ELISA, syncytium formation detected by microscopy. The different multiplicity of HIV-1/HSV-1 ratio was used. Results: The double viral HIV-1/HSV-1 infection was more cytopathic comparing with monoinfections. In mixed infection by the HIV-1/HSV-1 concentration of HIV-1 antigens and syncytium formations increased by 1,7 to 2,3 times in different cells in comparison with the culture infected with HIV-1 alone. The concentration of HSV-1 increased by 1,5-1,7 times, respectively. Administration of FPACA (1 microg/ml) protected cells: HIV-1/HSV-1 (1:1) – 80,1%; HIV-1/HSV-1 (1:4) – 57,2%; HIV-1/HSV-1 (1:8) – 46,3 %; HIV-1/HSV-1 (1:16) – 17,0%. Virus’s antigen levels were also reduced. Syncytium formation was totally inhibited in all cases of mixed infection. Conclusion: FPACA showed antiviral activity in case of mixed viral infection induced by Human Immunodeficiency Virus and Herpes Simplex Virus. The effect of viral inhibition increased with the multiplicity of HIV-1 in the inoculum. The mechanism of FPACA action is connected with the blocking of the virus particles adsorption to the cells and it could be suggested that it can have an antiviral activity against some other viruses too. Now FPACA could be considered as a potential drug for treatment of HIV disease complicated with opportunistic herpes viral infection.

Keywords: antiviral drug, human immunodeficiency virus (hiv), herpes simplex virus (hsv), mixed viral infection

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Authors: Marcin Debowski, Marcin Zielinski, Magdalena Zielinska, Paulina Rusanowska

Abstract:

The problem with digestate management is one of the most important factors influencing on the development and operation of biogas plant. Turbidity and bacterial contamination negatively affect the growth of algae, which can limit the use of the effluent in the production of algae biomass on a large scale. These problems can be overcome by cultivating of algae species resistant to environmental factors, such as Chlorella sp., Scenedesmus sp., or reducing load of organic compounds to prevent bacterial contamination. The effluent requires dilution and/or purification. One of the methods of effluent treatment is the use of a membrane technology such as microfiltration (MF), ultrafiltration (UF), nanofiltration (NF) and reverse osmosis (RO), depending on the membrane pore size and the cut off point. Membranes are a physical barrier to solids and particles larger than the size of the pores. MF membranes have the largest pores and are used to remove turbidity, suspensions, bacteria and some viruses. UF membranes remove also color, odor and organic compounds with high molecular weight. In treatment of wastewater or other waste streams, MF and UF can provide a sufficient degree of purification. NF membranes are used to remove natural organic matter from waters, water disinfection products and sulfates. RO membranes are applied to remove monovalent ions such as Na⁺ or K⁺. The effluent was used in UF for medium to cultivation of two microalgae: Chlorella sp. and Phaeodactylum tricornutum. Growth rates of Chlorella sp. and P. tricornutum were similar: 0.216 d⁻¹ and 0.200 d⁻¹ (Chlorella sp.); 0.128 d⁻¹ and 0.126 d⁻¹ (P. tricornutum), on synthetic medium and permeate from UF, respectively. The final biomass composition was also similar, regardless of the medium. Removal of nitrogen was 92% and 71% by Chlorella sp. and P. tricornutum, respectively. The fermentation effluents after UF and dilution were also used for cultivation of algae Scenedesmus sp. that is resistant to environmental conditions. The authors recommended the development of biorafinery based on the production of algae for the biogas production. There are examples of using a multi-stage membrane system to purify the liquid fraction from digestate. After the initial UF, RO is used to remove ammonium nitrogen and COD. To obtain a permeate with a concentration of ammonium nitrogen allowing to discharge it into the environment, it was necessary to apply three-stage RO. The composition of the permeate after two-stage RO was: COD 50–60 mg/dm³, dry solids 0 mg/dm³, ammonium nitrogen 300–320 mg/dm³, total nitrogen 320–340 mg/dm³, total phosphorus 53 mg/dm³. However compostion of permeate after three-stage RO was: COD < 5 mg/dm³, dry solids 0 mg/dm³, ammonium nitrogen 0 mg/dm³, total nitrogen 3.5 mg/dm³, total phosphorus < 0,05 mg/dm³. Last stage of RO might be replaced by ion exchange process. The negative aspect of membrane filtration systems is the fact that the permeate is about 50% of the introduced volume, the remainder is the retentate. The management of a retentate might involve recirculation to a biogas plant.

Keywords: digestate, membrane filtration, microalgae cultivation, Chlorella sp.

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Authors: Muhammad Farooq Baig, Saleem Qadeer

Abstract:

Background: The frequency of hepatitis C virus infection along with tuberculosis has not been widely investigated and very low statistics on rates of hepatitis C virus co-infection in tuberculosis patients. Hepatotoxicity is the major side effect of anti-tuberculosis therapy hepatitis HCVliver disease elevates the chances of hepatotoxicity up-to five folds. Objectives & Aim: To see the frequency of Hepatitis Cvirus infection amongst people with diagnosed Tuberculosis using gene X-pert technique. To evaluate the factors associated with HCVinfection in patients with MTBtuberculosis and to determine sensitivity and specificity of the tests. Study design: Comparative analytical study. Methodology: Three hundred and thirteen patients of tuberculosis diagnosed by Genexpert included while testing hepatitis C virus using immunochromotography rapid test technique, enzyme linked immunosorbent assay method and polymerase chain reaction test for confirmation. Results:Higher frequency of tuberculosis infection in males 57.8%, 42.5% between 20-39 years and 22% of hepatitis C virus infection in tuberculosis patients.The sensitivity of rapid test and enzyme-linked immunosorbent assay was 79% and 96% respectively while the specificity of rapid test and enzyme-linked immunosorbent assay was 91% and 99% respectively.

Keywords: Mycobactrium Tuberculosis, PC'R, Gene x pert, Hepatitis C virus

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Authors: Khabat Barkhordari, Ali Mohammad Arabzadeh

Abstract:

Hepatitis delta virus (HDV) is a RNA virus that needs the function of hepatitis B virus (HBV) for its propagation and assembly. Infection by HDV can occur spontaneously with HBV infection and cause acute hepatitis or develop as secondary infection in HBV suffering patients. Based on genome sequence analysis, HDV has several genotypes which show broad geographic and diverse clinical features. The aim of current study is determine the prevalence of hepatitis delta virus genotype in patients with positive HBsAg in Kerman province of Iran. This cross-sectional study a total of 400 patients with HBV infection attending the clinic center of Besat from 2012 to 2014 were included. We carried out ELISA to detect anti-HDV antibodies. Those testing positive were analyzed further for HDV-RNA and for genotyping using restriction fragment length polymorphism (RFLP) and RT-nested PCR- sequencing. Among 400 patients in this study, 67 cases (16.75 %) were containing anti-HDV antibody which we found HDV RNA in just 7 (1.75%) serum samples. Analysis of these 7 positive HDV showed that all of them have genotype I. According to current study the HDV prevalence in Kerman is higher than the reported prevalence of 6.6% for Iran as a whole and clade 1 (genotype 1) is the predominant clade of HDV in Kerman.

Keywords: genotyping, hepatitis delta virus, molecular epidemiology, Kerman, Iran

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Authors: Guler D. Uzel, Andrew G. Parker, Robert L. Mach, Adly Abd-Alla

Abstract:

Several tsetse fly species (Diptera: Glossinidae) in natural or colonized populations can be infected with the salivary gland hypertrophy virus (SGHV), a circular dsDNA virus (Hytrosaviridae). The virus infection is mainly asymptomatic but, in some species under certain conditions, the infection can produce salivary gland hypertrophy (SGH) symptoms. In the laboratory colonized tsetse, flies with SGH have reduced fertility, which negatively affects colony performance. Therefore, a high prevalence of SGH in insect mass rearing represents a major challenge for tsetse control using the sterile insect technique. The main objective of this study is to analyze the impact of Glossina pallidipes SGHV infection in various tsetse species on mortality and productivity and its impact on the symbiotic bacteria. Hypertropied salivary glands (SG) were collected from G. pallidipes into phosphate buffered saline (PBS) to prepare suspension; 2 µl aliquots were injected into adults of several tsetse species (G. pallidipes (Gp), G. p. gambiensis (Gpg), G. brevipalpis (Gb), G. morsitans morsitans (Gmm), G. morsitans centralis (Gmc) and G. fuscipes (Gf)) and the change in virus and symbiont titers were analyzed using qPCR. The development of SGH in the F1 was detected by dissection 10 days after emergence and virus infection was confirmed by PCR. The impact of virus infection on fly mortality and productivity was recorded. 2 µl aliquots were also injected into 3rd instar larvae of the different species and the adult SGs assayed by PCR for virus. Virus positive SGs from each species were homogenized in PBS and pooled within species for injection into larvae of the same species. Flies injected with PBS were used as control. Injecting teneral flies with SGHV caused increasing virus titer over time in all species but no SGH was detected. Dissection of the F1 also showed no development of SGH except in Gp (the homologous host). Injection of SGHV did not have any impact on the prevalence of the tsetse symbionts, but an increase in Sodalis titer was observed correlated with fly age regardless of virus infection. The virus infection had a negative impact on productivity and mortality. SGHV injection into larvae of the different species produced SGHV infected glands in the adults determined by PCR with a rate of 60%, 27%, 16%, 7% and 7% for Gp, Gf, Gpg, Gmm and Gmc, respectively. Virus positive SGs observed in the heterologous species were smaller than SGH found in Gp. No virus positive SG was detected by PCR in Gb and no SGH was observed in any adults except in Gp. Injecting virus suspension from the virus positive SGs into conspecific larvae did not produce any adults with infected SGs (except in Gp). SGHV can infect all tested tsetse species. Although the virus can infect and increase in titer in other tsetse species and affect fly mortality and productivity, no vertical virus transmission was observed in other tsetse species with might indicate a transmission barrier in these species, and virus collected from flies injected as larvae was not infective by injection.

Keywords: DNA viruses, glossina, hytrosaviridae, symbiotic bacteria, tsetse

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