5^th World Congress on Virology December 07-09, 2015 Atlanta, USA

Shaheen has completed his PhD at the age of 31 years in the field of virology from Al-Azhar University. Also, Dr. Shaheen work as researcher of molecular virology at National Research Center, Dokki, Egypt. Shaheen has eight years experience in the field of virology. To date he has published nine papers in reputed journals and three abstracts in in international conferences, most of these publications he is the first and corresponding author. Currently, Dr. Shaheen started postdoctoral training in virology field at molecular medicine department, Padova university, Italy.

Coxsackie virus B3 (CVB3) represents current major threats to public health and considers as an important viral pathogen related to viral myocarditis. We determined the antiviral properties of five extracts (methanol, chloroform, ethyl acetate, n-butanol, and aqueous ) from Cassia alata leaves in vitro. The most potent extract was selected to be tested in vivo. In vitro, the cytotoxicity effect of the extracts on GMK cells was conducted by MTT colorimetric assay. The antiviral activity of the extracts was determined in three different ways (virucidal, pre-treatment, and post-treatment) by MTT and 50% tissue culture infectious dose (TCID50) methods. In vivo, after toxicity determination, the antiviral activity of the selected extract using two safe doses was evaluated based on determination of the morbidity, mortality, heart to body weight ratio (HW/BW), activities of LDH, AST, and CK enzymes, virus titers, and necrosis in heart tissues in infected mice with CVB3. Our results demonstrated that the all extracts of C. alata showed antiviral activity against CVB3 in vitro with TI ranged from 0.2 to 12.2 and reduction in virus titers ranged from 0 log10 to 2.5 log10 where the aqueous extract was the most effective against CVB3 infection in vitro. In vivo, the aqueous extract was found to be safe at 100 mg/kg body weight and therefor for antiviral evaluation we used 100 and 50 mg/kg body weight as safe doses. Our results suggested that the aqueous extract of C. alata significantly reduced the morbidity, mortality, HW/BW, virus titers, necrosis and mononuclear cell infiltration of heart tissues at the both dosages. Also the extract showed the ability to maintain levels of LDH, AST, and CK enzymes at normal level in the treated infected mice compared with those untreated infected mice. This result suggested that the aqueous extract of C. alata may represent a potential antiviral agent to treatment CVB3 myocarditis.

I’am Hanen Boukoum Dr on Biology. I Have completed my PhD this year 2015. my research work was concentrated mainly on the study of viruses. My thesis was done in the laboratory of transmissible diseases and biologically active substances in the Faculty of Pharmacy of Monastir, Tunisia and a part of my work was made in the laboratory of Virology in Saint Eloi hospital Montpellier in France. Currently I have five publications in the JMV.

BK polyomavirus (BKPyV) is the most common pathogen in renal transplant recipients and BKPyV associated nephropathy (BKPyVAN) is one of the most significant causes of graft dysfunction and loss in renal transplant recipients. The aim of this study is to find if there is an association between the BKPyV genotypes and the BKPyVAN. A total of 72 renal transplant recipients were studied. BKPyV were detected and quantified by real-time PCR in urine and plasma. Isolates from positive urine samples were genotyped and a phylogenetic analysis was performed. In the studied population, two from the 34 BKPyV infected patients developed BKPyVAN and both have subtype I/ subgroup Ib-2. The most other patients clustered with genotype I/ subgroup Ib-2 present in 76.5% of the detected isolates. Subtype II, Subtype IV and Subtype I/subgroup Ib-1 were found in 17.5%, 3% and 3%, respectively. This study suggests that there is not a clear relationship between viral genotypes and BKPyVAN; since the majority of the asymptomatic infected patients had the subtype I/subgroup Ib-2.

Uladzimir Karniychuk has obtained a DVM degree with Honours from Vitebsk State Academy of Veterinary Medicine, Belarus. He has completed his PhD in the Laboratory of Virology, Ghent University, Belgium. At present, he is a postdoctoral researcher in University Health Network, Canada. He has published 15 papers in well-recognized international journals and obtained several highly competitive international and national fellowships (three PhD grants, CIHR Postdoctoral Fellowship) and awards (Toronto General Research Institute Postdoctoral Fellowship Award and six young scientist travel/poster awards).

Most patients with severe influenza disease seek hospital care many days after initial onset of symptoms, limiting the effective use of antivirals. The main goal in the present study was to compare therapeutic effects of delayed intrapulmonary and intramuscular delivery of Peramivir on severe infection outcome. To recapitulate late stage, lower respiratory tract infection, aged ferrets were inoculated intrapulmonary with 106 EID50 influenza A/California/07/2009 H1N1. Peramivir was delivered at 3 and 5 dpi intrapulmonary (PP) or intramuscularly (PM). A group of animals was administrated intrapulmonary with only a vehicle (saline) for control purposes (VP). All animals in the VP group were euthanized due to severity of the diseases at 8 dpi. Survival in the PP group (75%) was 25% higher than in the PM group. In comparison to the control VP group, intrapulmonary and intramuscular Peramivir administration efficiently reduced viral titers in nasal washes. At 5 dpi, viral titers in lungs from the PP group were lower than in samples from other groups (PP – mean 3.37 TCID50/ml; PM – mean 4.55 TCID50/ml; VP – mean 6.44 TCID50/ml). Reduced area of lung surfaces with pathology were observed in the PP group (PP – mean 19%; PM – mean 40%; VP – mean 72%). Microscopic lesions associated with severe disruption of the lung architecture were found only in ferrets from the VP group and one ferret from the PM group. To summarize, delayed intrapulmonary Peramivir administration in aged ferrets has benefits to recovery from severe influenza infection in comparison to delayed intramuscular Peramivir administration.

Tatyana Kushnareva is the leading researcher in Hantaviral infections laboratory in Institute of Epidemiology and Microbiology, Vladivostok, Russia. She is the executive of the section of basic researches on disclosure of hantavirus circulation mechanisms in the nature and regularities of natural foci functioning in various landscape, development and improvement of forecasting methods of HFRS incidence on endemic territories. She is directly involved in carrying out the epizootological researches in the field conditions in natural foci. Within the last five years she published 48 works: 13 in reviewed magazines, 30 in periodic scientific publications and Conference materials, 4 Databases, Methodical recommendations.

Persistence of Pathogenic for Man Hantaviruses in Apodemus Mice Populations (E-BABE): Objective: Complex monitoring of dynamics of epizootic and epidemic processes on HFRS endemic areas in Primorye Region of Far East Russia with the purpose of evaluation persistence of Hantaan (genovariant FE) and Amur hantaviruses and detection of periods of acute infection in theirs rodent-reservoir populations – Apodemus agrarius and A. peninsulae accordingly – were performed. Methods: Epizootological monitoring was carried out in the forest-steppe landscape favourable for A.agrarius and in the cedar-oak forests optimum for A.peninsulae (2001-2013). Criteria of acute infection were detection of viral antigen/RNA in the lungs and organs of secretion/excretion and low avidity antibody in blood of rodents using ELISA, RT-PCR and IFA. Cases of HFRS diseases were summed in every epidemic season. Results: The specific antigen/RNA in eliminative organs were detected of 2.0% A.agrarius and 1.6% A.peninsulae in period of epizootic activity increase and 11.8% and 9.6% respectively during of high epizootic activity. Period of low epizootic activity in different Apodemus mice populations was associated with 0.2% A.agrarius and 0.7% A.peninsulae with acute infection. In year of high activity of epizootic process seasonal dynamic of hantaviral infection in A.agrarius populations was characterized by increase of infected mice from spring (3.5%) to autumn (13.4%) and on the contrary by decrease in A.peninsulae populations (18.1% and 4.0% respectively). Animals with hantaviral antigen in eliminative organs were mainly at 3-5 months old and 67.4% from them had low avidity IgG antibodies. Annual variations of epidemic activity of different HFRS natural foci directly correlated with numbers of carriers of acute hantaviral infection in A.agrarius and A.peninsulae populations. Conclusion: Outbreaks of HFRS on Primorye Territory were observed at the same year of high epizootic activity in hantaviral infection natural foci. Thereby we can conclude that periods of acute infection in Apodemus mice populations cause increase of HFRS morbidity.

S M Rashed-ul Islam is a virologist and researcher, working at the Department of Virology of Bangabandhu Sheikh Mujib Medical University (BSMMU). He has obtained his M. Phil in Medical Virology in 2012 from Bangabandhu Sheikh Mujib Medical University (BSMMU). He is actively involved in the academic, research and laboratory work and also attended several conferences at both national and international levels. Besides virology, Dr. Islam has special interest in many Public Health focuses with his keen concern on emerging viral diseases (Nipah, Influenza), HIV/AIDS, Hepatitis B&C, universal health coverage, double burden diseases, etc. Until now, Dr. Islam has published eight articles in peer reviewed national and international journals.

The world is now under human pandemic threat by avian influenza viruses. As the human, animal and the environment interact closely from the dawn of the civilization, human health is tremendously influenced by animal health and their health issues. Like other countries, Bangladesh has also reported human cases of avian influenza subtype H1N1, H3, H5N1, and H9N2 across the country. Influenza surveillance is being conducted since 2007 in 12 hospitals covering administrative divisions of the country in collaboration with IEDCR and ICDDR’B. National Influenza Surveillance, Bangladesh (NISB) started operating from April 2010 in 14 district hospitals under direct supervision of IEDCR. The National Rapid Response Team (NRRT) is under 7/24 alert and conducting investigation of reported cases of any suspected Influenza. IEDCR is also responsible for overall coordination and subsequent report preparation periodically. There is a “National Avian Influenza and Human Pandemic Influenza Preparedness and Response Plan” in Bangladesh to prevent and control avian and human pandemic influenza. The preparedness plan encompasses strategy development including planning & coordination, surveillance, prevention & control, risk communication, and operational research. The plan offers different strategies for different scenarios (whether Bangladesh is affected or not) involving both human and avian health through multi-sectoral collaborative approach. The best way to prevent infection with avian influenza viruses is to avoid sources of exposure i.e. direct or close contact with infected poultry. Frequent hand wash with soap-water, maintenance of hygiene and infection control measures can prevent influenza virus infection.

S M Rashed-ul Islam is a virologist and researcher, working at the Department of Virology of Bangabandhu Sheikh Mujib Medical University (BSMMU). He has obtained his M. Phil in Medical Virology in 2012 from Bangabandhu Sheikh Mujib Medical University (BSMMU). He is actively involved in the academic, research and laboratory work and also attended several conferences at both national and international levels. Besides virology, Dr. Islam has special interest in many Public Health focuses with his keen concern on emerging viral diseases (Nipah, Influenza), HIV/AIDS, Hepatitis B&C, universal health coverage, double burden diseases, etc. Until now, Dr. Islam has published eight articles in peer reviewed national and international journals.

The management of HIV/AIDS in Bangladesh has been greatly transformed because of widespread availability of Anti-retroviral Therapy (ART). The present study was undertaken to determine the clinico-epidemiological and immune monitoring among the Bangladeshi HIV/AIDS patients. A cross-sectional study was carried out among 100 HIV/AIDS patients divided into Therapy naïve (Group I; n=33), Symptomatic Therapy naïve (Group II; n=33) and ART receivers >1 year (Group III; n=34) at the Department of Virology of Bangabandhu Sheikh Mujib Medical University (BSMMU) between January and December of 2011. Patients were monitored during their clinical follow-ups and CD4 & CD8 T-lymphocyte were analyzed using Flowcytometer (BD FACS count, USA). Of the 100 HIV/AIDS patients, 53 (53%) were males, 43 (43%) were females and 4 (4%) were transgenders (TGs). The age of the study population ranged from 19 to 60 years (mean ±SD; 33.53±9.07 years) with predominant age group between 30-40 years. The most frequent (84%) mode of transmission was heterosexual route. 76% of Group I and 91% of Group III were asymptomatic as per their clinical presentation. Among Group II, majority (58%) of individuals were presented with a combination of fever, diarrhea, weight loss and oral infection. As per WHO Clinical staging’s, 81% of Group I were belong to Stage 1, whereas, in Group II, 42% and 36% were belong to Stage 3 and Stage 4 respectively. CD4 and CD8 T-lymphocyte were 591±217 and 1266±433 cells/μl respectively in Group I. Among Group II, CD4 T-lymphocyte were found reduced significantly to 155±127 cell/μl (P<0.05) while CD8 T-lymphocyte were 758±564 cell/μl (P<0.05) when compared with Group I. Among Group III, CD4 and CD8 T-lymphocyte count were 588±228 and 1104±417 cells/μl respectively with no significant difference with Group I suggesting the intake of ART has prominently bring back the immune function to near normal. All the symptomatic therapy naïves with CD4 T-lymphocyte count <350 cells/μl were subsequently counseled for ART program. Timely initiation of ART and clinical/immune follow-up ensures the success of the ongoing national scale up of the ART Program in Bangladesh.

Sareh Zhand is a PhD candidate of microbiology at Shahid Beheshti University of Iran. She specially work on Hepatitis B virus gene mutations and has published more than 5 papers in reputed journals and has more than 18 abstract presented in international congresses and she is the winner of the first Prize for oral Presentation from the 4th International Hepatitis Congress and winner of the First Prize for Poster Presentation from 6th International Hepatitis Congress.

Introduction: Hepatitis B virus (HBV) represents a major health problem worldwide, developing cirrhosis and hepatocellular carcinoma (HCC) in some cases. Mutations in the HBV surface (S) and Polymerase regions has been observed frequently associated with the virus replication, Vaccine and Immune Escape mutations, Drug resistance and clinical outcomes ,respectively. This study aimed to investigate mutation in these regions of chronic HBV patients (CHB) in Golestan province, Northeast of Iran. \r\n\r\nMaterials and Methods:\r\nThis cross sectional study is done on 65 CHB (Chronic HBV) patients (HBsAg positive for more than 6 months), which were under Lamivudine treatment. HBV-DNA was extracted and PCR was performed using specific primers for four regions including S gene and Polymerase region. Positive PCR products were subjected to automated sequencing. Alignment was applied using reference sequence from Gene Bank database with AB033559 accession number. \r\n\r\nResults: \r\nResults showed that 11% of our patients had mutations in the ‘‘a’’ determinant of S gene, one case with G145R mutation was detected, which is called \"Escape Mutation\" and is reported for the first time from IRAN. Mutations at the YMDD and FLLAQ motifs in the polymerase gene of HBV detected in 12 of 65 patients (18.46%) which were under treatment with Lamivudine. \r\n\r\nConclusion:\r\nMutations in Polymerase and S regions of the HBV genomes in this Iranian CHB population is very important to predict the outcome of infection, The administration of antiretroviral drugs, especially it is very useful for predicting the efficiency of HBV vaccines.\r\n

Thalita Ferreira has graduated in Pharmacy at the University of Brasília. During graduation she\r\nintegrated the research group led by Enrique Argañaraz in the Molecular Virology Laboratory.\r\nCurrently is a doctoral student at the University of Campinas integrating the research group in Drug\r\nDiscovery led by Lúcio Holanda FreitasJúnior.

Large variation exists in susceptibility to infection with Human Immunodeficiency Virus\r\nType 1 (HIV), and disease progression. These observations demonstrate a role for antiretroviral host\r\nfactors. Several reports describe a1antitrypsin\r\n(A1AT), the most abundant circulating serine\r\nprotease inhibitor, as a potent suppressor of HIV infection and replication. We identified the normal\r\n(M) and most common deficiencyassociated\r\n(S and Z) isoforms of the A1AT gene in patients\r\ninfected with HIV from four multicenter cohorts. The level of disease progression in the patients\r\nwas characterized and the patients were grouped into as elite controllers (EC), longterm\r\nnonprogressors\r\n(LTNP), or progressors (Prog). No significant difference in the distribution of\r\nA1AT alleles was observed in the EC, LTNP, or Prog groups. However, significantly increased\r\nprevalence of the A1AT deficiencyassociated\r\nS allele was observed in HIVinfected\r\npatients\r\ncompared to the prevalence of S A1AT in the general population. These results suggest that\r\ndeficiency in A1AT may be a risk factor for acquisition of HIV infection, but physiological A1AT\r\nconcentrations do not affect disease progression after infection occurs.

Ivana has completed her PhD at the age of 26 years from University of South Bohemia in Czech Republic and postdoctoral studies from Vienna University in Austria as well as La Jolla Institute for Allergy and Immunology in California. She has a junior project leader position at the Institute of Virology at the Slovak Academy of Sciences and she is proudly using a status of Marie Curie Fellow. She has 8 publications and more than 60 citations in reputed journals.

The immune system has evolved to protect against the many pathogens that are encountered throughout the lifetime of an individual. In turn, the selective pressure that is exerted by the immune system has shaped pathogen evolution. This co-evolutionary relationship between host and pathogen is particularly clear for viruses that establish persistent infections, such as human herpesviruses. Human cytomegalovirus (HCMV, a β-herpesvirus), is a large double-stranded DNA virus that causes a lifelong, persistent/latent infection in ~50-80% of the US population. While HCMV infection is largely asymptomatic in healthy persons, it can induce serious disease in those with naïve or compromised immunity, and the high incidence of congenital infection has spurred a strong initiative for vaccine development. Primary clinical isolates carry at least 19 additional genes within the UL/b’ genomic region that have been lost in several commonly used HCMV strains, with several of them targeting signaling by the TNF superfamily. Here, we show that the immunomodulatory function of HCMV UL141 is associated with its ability to bind diverse proteins, while utilizing at least two distinct binding sites to selectively engage TRAIL DRs or CD155. Binding studies revealed high affinity interaction of UL141 with both TRAIL-R2 and CD155 and low affinity binding to TRAIL-R1. We determined the crystal structure, which revealed that UL141 forms a homodimer that engages two TRAIL-R2 monomers 90° apart to form a heterotetrameric complex. The breadth of UL141-mediated effects indicates that HCMV has evolved sophisticated strategies to evade the immune system by modulating multiple effector pathways. Reference: Nemcovicova, et al. 2013 and 2014.

Ana Godinho-Santos has a degree in Microbiology and a master degree in Applied Microbiology. She is currently a doctoral candidate at Faculty of Pharmacy - University of Lisbon, expected to complete the PhD degree requirements by March 2016.

Human Immunodeficiency Virus type 1 (HIV-1) relies on the host cell machinery to complete its life cycle. Several helper factors have been identified in expectation that a better understanding of host-HIV interactions would lead to novel antiviral targets. As the characterization for many of these proteins is still limited, we aimed to depict the contribution of CIB2, previously identified as helper factor, as well as the potential contribution of its homolog, CIB1. Knockdown of both CIB1 and CIB2 in shRNA-transduced cell populations strongly impaired viral replication in target cells, recognizing these proteins as non-redundant HIV-1 helper factors. Also, a single-round cycle assay demonstrated that normal levels of CIB1 and CIB2 are required for HIV-1 envelope-mediated process. In fact, both X4 and R5 viral strains seem to be dependent on CIB1 and CIB2 expression levels, since infectivity of X4- and R5-enveloped particles was affected. Moreover, CIB1- and CIB2-knockdown populations displayed reduced viral fusion efficiency when compared to untransduced population, indicating virus-free entry impairment. Furthermore, virus-transfer through cell-cell contacts was decreased after co-culture of infected donor cells with CIB1- and CIB2-knockdown target cells. Flow cytometry showed that surface expression of some key players of both routes of viral entry was altered in CIBs-depleted populations, namely co-receptor CXCR4 and integrin α4β7. Taken together, these studies revealed for the first time that CIB1 is a HIV-1 helper factor along with CIB2, and suggest that both CIB1 and CIB2 facilitate HIV-1 entry in natural target cells possibly through modulation of CXCR4 and α4β7.

Safder is a PhD candidate of Microbiology at the University of Kansas Medical Center Kansas City, USA. He has obtained his masters degree in Biotechnology from the University of Kashmir, India. His work is much focused on replication, transcription and protein translation of different viruses like hantavirus and parvovirus. He has published several papers in various reputed journals.

Hantaviruses are the causative agents of hemorrhagic fever with renal syndrome (HFRS) and hantavirus cardio-pulmonary syndrome (HCPS) in humans. Hantaviruses, Bunyaviridae family members, are tripartite negative strand RNA viruses. The three segments of genome, L, M and S encode RNA dependent RNA polymerase (RdRp), glycoproteins (G1 & G2) and nucleocapsid (N) protein, respectively. Nucleocapsid protein (N) plays diverse roles during hantavirus infection. Primarily, N protein is involved in the encapsidation and packaging of viral genome. Hantavirus N protein helps in the preferential translation of viral mRNAs by specifically binding to the 5’UTR of viral mRNAs and recruiting translation initiation machinery. N protein also binds 5’ Cap and facilitates translation initiation by acting as eIF4F surrogate. We have recently found that N protein interacts with 40S ribosomal subunit via ribosomal protein S19 (RPS19). Using deletion mutagenesis approach, we identified RPS19 binding domain at the N terminus of N. Further, fusing RPS19 binding domain with GFP was sufficient to pull down RPS19 protein. N-mutant deficient in RPS19 binding didn’t augment reporter mRNA translation both in vivo as well as in vitro. Upon the inhibition of cap dependent translation, the mutant didn’t support the translation of reporter mRNA (GFP) as the wt-N did. On characterizing the mutant, it was observed that no other known functions of N protein were compromised, like any major structural change, trimerization, viral RNA panhandle and mRNA cap binding. Our studies revealed another new target (N protein and RPS19 interaction) that can be subjected to therapeutic intervention for hantavirus infection.

Sakshi Sharma is currently working as a UGC Junior Research Fellow/PhD Scholar from DEI. Her study basically includes screening, identification, transmission, purification and characterization of mycoviruses. She got best paper presentation/young system scientist award in the conference, Paritantra (System Society of India) in 2013. She received International Travel Grant Award by Science and Engineering Research Board (SERB), under the Department of Science and Technology (DST), New Delhi as a Young Scientist in 2011 to present an oral paper in International Conference, Autoimmunity Congress Asia (ACA), Singapore. She has papers published in Indian and International SCI journals.

Candida albicans is widespread yeast; the infections can be short lived, superficial skin irritations to devastating, fatal systemic diseases. The confirmation of viruses in fungi commonly called Mycoviruses or VLP’s. In present study, this human pathogenic fungus was screened for the presence of virus like particles (VLP’s). Negative staining and transmission electron microscopy positively indexed this fungus for the presence of VLP’s. The concentration of VLP’s is good and the particles are isometric in shape with 40 to 66 nm in size. Further, treated with cycloheximide (50 µg/ml) revealed the complete elimination of VLP’s. Extraction and purification of nucleic acid from CF 11 column chromatography gave positive results whereas extraction from SDS phenol was found to be unsuccessful. For establishing the nature of nucleic acid, RNAse and DNAse treatment was followed. DNAse treatment and high salt (1SSC) did not revealed any band in Candida albicans whereas low salt (0.01SSC) RNAse test of Candida albicans reveals a band which concluded that the nature of nucleic acid is dsRNA. Subsequently, thermo melting confirms the nature of nucleic acid. Current finding may help in understanding and using the dsRNA mycovirus in fungal bio-control, fungal infections and also magnifies our knowledge of virus ecosystem and development.