E-mail: secretary@virologycongress.com | USA : +1-646-828-7579, UK : +44-203-695-1242 | August 21-23, 2017, Barcelona, Spain  


Anusha Fernando
Medical Research Institute, Colombo, SriLanka
Title: An outbreak of concurrent infections with dengue virus serotypes in Kinniya, Trincomalee ,SriLanka

Biography: Anusha Fernando is from Medical Research Institute, Colombo, SriLanka

Abstract: Introduction Co-circulation of multiple dengue virus serotypes has been reported from many parts of the world including Sri Lanka.However, concurrent infection with more than one serotype of dengue viruses in the same individual is rarely documented. An outbreak of dengue hemorrhagic fever/dengue shock syndrome occurred in most parts of the country in 2017, though significant mortalityreportedonly from Kinniya, in Trincomalee District. We investigated the dengue out break with serological and molecular test methods. Objective To determine the acute dengue infection and describe the dengue virus serotypes in the Kinniya outbreak, in 2017. Methodology The study analyzed total22 symptomatic serum samples which were sent to Medical Research Institute, for routine testing for dengue virus infection during third week of March, 2017. Samples were from patients within 7 days of fever and presented to Base Hospital Kinniya. The received samples have been tested for dengue virus infection, usingmultiplex typingrRT-PCR assay (Waggoner etal 2012),NS1 rapidimmunochromatographicassay, andDengue IgM ELISA (SD Korea). All the samples were tested with multiplex typingrRT-PCR assay, and Dengue IgM ELISA assay. Only 13 samples tested with dengue NS1 immunochromatographic assay. Results All 22(100%) samples gave evidence for positive dengue virus infection at least through one of the above mentioned assays. Of the tested samples, 86%, 68% and 38% were positive with typing rRT-PCR, Dengue IgM ELISA and NS1 rapid immunochromatographic assay respectively.All rRT-PCR assay positive samples showed up DENV-2,followed by DENV-4(73%) and DENV-1(5%). Concurrent infection identified among 79% of therRT-PCR assaypositive samples. 74%samples were infected with both DENV-2 and DENV-4 and 5 % co-infected with DENV-2 and DENV-1. Single serotype 2 infection was identified in 21% samples. Conclusions The results convincedKinniya outbreak is an acute dengue virus infection. It also revealed thatDENV-2 dominated the outbreak whereas DENV-2 and DENV-4 lead to concurrent infection. The dengue virus type 1 responsible only for a lesser extent for this outbreak.

Janaki Abeynayake
Medical Research Institute, Colombo, SriLanka
Title: A viral etiology and clinico-epidemiological profile of clinically suspected acute viral encephalitis

Biography: Janaki Abeynayake is from Medical Research Institute, Colombo, SriLanka

Abstract: Introduction: Acute viral encephalitis isa medical emergency which could lead to considerable morbidityand mortality. The spectrum of brain involvement,with varying severity and the outcome of the disease depend on the virus. Geography is a major determinant for encephalitis caused by vector-borne viruses. This retrospective study will focus on Flaviviral etiology, diagnosis, geography and clinical profile. Objective:To describe Flaviviral etiology and clinico-epidemiological profile of acute viral encephalitis Methodology:This study included total of 381clinically Japanese Encephalitis (JE)suspected acute encephalitis patient samples from December 2016 throughFebruary 2017. The CSF samples received from both children and adults across the country were first testedforJE usingJE IgM Capture Enzyme Linked Immunosorbent(ELISA) assay (NIV Pune). The positive samples for JE IgMwere tested with Anti Dengue IgMcapture ELISA (SD Korea) assay and to rule-out dengue with dengue real time typing PCR assay.Patients were diagnosed with dengue if they were positive for dengue RNA.Only detectable JE IgMwas judged to have JE infection, while detectable JE IgM and dengue IgM but not dengue RNA, were identified as Flavivirus infection. The clinico-epidemiological profiles sent along with the sample were analyzedondiagnosed cases. Results:A total of 381 CSF samples weretested and 23(6%)were positive for JEIgM.Of the JE positives,08(35%) were positive for dengue IgM and only 04(17%) positive for dengue RNA.JE only was positive in 11(48%) patients.Dengue and JE serology both positive in8(35%) while 4(17%) were confirmed with dengue RNA. Allcases experienced fever. Altered consciousness and seizures observed in 90%.It was noted that 60% of JE reported from Sabaragamuwaprovince andyoung adults and elderly were affected most. Conclusions:JE and dengue identified as contributing viral etiology. Even though dengue is not primarily neurotropic, the study convinced the importance of considering dengue encephalitis in appropriate situations. Findings appreciated common clinical profile of acute encephalitis to intervene timely.Data explained the geography ofhigh JE activity and the most affected age group tostrengthen the preventive/control measures.

Yen-Hung Chow
National Health Research Institutes, Zhunan ,Taiwan.
Title: Mutations in VP1 protein determinates enterovirus 71 virulence

Biography: Dr. Yen-Hung Chow is an associate Principal Investigator at National Health Research Institutes, Taiwan since 2012. He also cross-appointed as an associate Professor at the China Medical University, Taiwan from 2015 to present. His research focuses on development of a mucosal vaccines against the respiratory syncytial virus (RSV) utilizing the following technologies and strategies. He created an emterovirus infectious mice model for EV71 infection and developed a multivalent vaccine against HFMD (Foot-and-Mouth Disease). He has published over 47 peer-review articles and awarded 10 patents. He serves as reviewers for several international journals and executive members in number of scientific societies.

Abstract: Enterovirus 71 (EV71) is a major caustic for hand, foot and mouth disease (HFMD) and severe CNS complications associated with high mortality. Current genotype C2 of EV71 propagated in african green monkey kidney (Vero; EV71-V) or sub-passaged into human rhabdomyosarcoma (RD; EV71-R) cells as pathogen sources was used. Interestingly, one or two times-passaged EV71-R produced from the original isolates, EV71-V, elicited differential virulence in transgenic mice which express the main receptor of EV71, human scavenger receptor class B 2 (hSCARB2-Tg). Challenge of hSCARB2-Tg mice with EV71 showed that EV71-V behaved more virulence than EV71-R; in which the Tg-mice received lethal amounts of EV71-V resulted in 100% death, compared to all mice were alive while received EV71-R. The severe limb paralysis as well as muscle tissue damage that correlated with tissue viral burdens and proinflammatory cytokine secretions in EV71-V-mice, but observed mild symptoms associated with few virions and cytokines in EV71-R-mice. Consensus sequence analysis revealed that EV71-V aligned the amino acid sequence with EV71-R, rapidly acquired E145G, V146I, and S241L in VP1 and nucleotide T to C in 5’-UTR 494 substitutions. These mutations appear to have accumulation in response to growth adaptation to RD cells. EV71-R (145G) elicited higher binding affinity to another EV71 receptor of human P-selectin glycoprotein ligand-1 (hPSGL-1), compared to a non-hPSGL-1 binding strain of EV71-V (145E). However, EV71 have no difference in binding with hSCARB2. As expected correlation to the affinity of receptor binding, hPSGL-1-expressing L929 cell is susceptible to EV71-R but resistant to EV71-V, whereas EV71-V and EV-R show the similar infectivity in hSCARB2-expressing 3T3 cell. Higher yield of EV71-R than EV71-V in RD, PSGL-1-, and SCARB2-produced cells was observed. The residues substitutions predicted by molecular modeling indicate that these mutations in VP1 are corresponding to influence EV71’s engagement with PSGL-1 and in vivo virulence. Our results demonstrate substitution of some residues in hot spots of VP1 during propagation of EV71 in RD cell might occur and then affecting viral infection and pathogenesis in the host.

Maha AlKhazindar
Faculty of Science, Cairo University, Egypt
Title: Isolation and characterization of two bacteriophages infecting a strain of Kosakonia sacchari causing potato soft rot disease

Biography: Maha AlKhazindar is a Assistant Professor of Botany/Microbiology, Botany Department, Faculty of Science, Cairo University,Egypt. Research activities are focused on plant viruses and bacteriophages. Molecular and biotechnological approaches for the characterization and controlling of viral and bacterial diseases infecting crop plants. New and emerging viruses affecting nursery and landscape industry tospoviruses, phytoplasma. Epidemiology, and molecular studies on economically important plants infected with viruses and bacteria.

Abstract: Soil samples were collected from potato rhizosphere from a potato cultivated area in Giza, Egypt. Two bacteriophages were isolated on an endophytic bacterium causing potato soft rot (that was isolated and identified in our lab). The phages were purified and characterized using Dilution End Point (DEP), Longevity In Vitro (LIV), Thermal Inactivation Point (TIP), pH effect and host range. Based on genome characterization and electron microscopy, the two phages were named as vB_KsaM-C1 belonging to Myoviridae and vB_KsaO-C2 belonging to Microviridae according to Kropinski et al. [1]. Both phages sustained their activity up to 20 months with a remarkable decrease in phage titre. In addition both recorded the same TIP (65oC), and showed the optimum activity at neutral pH, However the DEP of vB_KsaM-C1 was at 10-7 while that for phage vB_KsaO-C2 was at 10-9. A small scale application of the two isolated phages on infected potato tubers discs successfully inhibited the soft rot disease.The two isolated bacteriophages showed positive results in inhibiting potato soft rot caused by the isolated host bacterium (Kosakonia sacchari) and controlled other strains of bacteria causing potato soft rot. Significance of study The two isolated phages can be used as a biocontrol agents in protecting potato tubers from soft rot disease either in the field or storage.

Sara Lafar
University of Hassan II of Casablanca, Morocco.
Title: Capripoxvirus disease: Isolation, molecular detection and epidemiological analysis of sheep pox in Morocco.

Biography: Sara LAFAR is a PhD student in “Biotechnology: Microbiology and Molecular Biology” at the Faculty of Sciences and Technologies of Mohammedia, University Hassan II of Casablanca (Morocco), working in collaboration with the pharmaceutical company “BIOPHARMA” in Rabat (Morocco).

Abstract: Sheep pox is a highly contagious disease caused by Capripoxvirus genus which belongs to Poxviridae family. The clinical signs include fever, skin lesions, lung nodules and death. The pathology has a devasting effect on livestock industry and inflicts substantial losses in terms of reduced productivity and lower quality of meat, wool and leather. The objectives of work were to realize an epidemiological analysis to understand the spatiotemporal evolution, to highlight the key factors involved in the spread of sheep pox and to confirm by isolation and RT-PCR the presence of virus in some regions in Morocco. The epidemiological analysis showed that sheep pox appears confined to the central and eastern regions where a very intensive sheep breeding activity is taking place with an average of 350 cases per year. The incidence varies depending on provinces and there was a positive correlation between the endemicity and the significant factor of rural market (P = 0,006). The annual average morbidity and mortality rates were 2,96% and 0,71% respectively and the intensity of the clinical signs were more pronounced in lambs, light-skinned races and even more in females than males. The molecular analysis has confirmed the presence of the virus and Ct values were between 19,39 and 42,5. The causal strain was isolated from lung nodules and the infective titre was 1,48.106 DICT50. This analysis allowed us to provide interesting informations to enhance the effectiveness of control and to implement an adequate strategy to fight against sheep pox in endemic countries.

Marcin Chodkowski
Warsaw University of Life Science Poland
Title: NucleoCounter NC-3000 – an efficient technique for the comprehensive assessment of cultured neurons state during EHV-1 infection

Biography: MarcinChodkowski is a PhD candidate in the Division of the Veterinary Medicine in Warsaw University of Life Sciences. His main research topic is:“Changes in the mitochondrial network in primary murine neurons infected with EHV-1”. He is interested in molecular virology,neurovirology and novel treatment approaches using viruses e.g. oncolytic viruses.

Abstract: The NucleoCounter NC-3000, a portable high-speed cell counting device based on the principle of fluorescence microscopy, provides the alternative method for standard flow cytometry. This system enables to perform automated cell counting and high precision analyses of eukaryotic cells during virus infections. The main objective of our study was to apply an efficient technique for assessment of the primary murine neurons culture infected with either neuropathogenic or non-neuropathogenic strains of Equine Herpesvirus type 1 (EHV-1). With NucleoCounter NC-3000 we have examined the cells viability, phase of the cell cycle and mitochondrial potential in cultured neurons infected with EHV-1. The apoptosis wasinvestigated, since we have already found that EHV-1 could influence this process [1]. Virus replication and transmission depends upon the existing molecular machinery of infected cell. Moreover, the latent herpesvirus infections prevent the cell death. lead to the cell death prevention. Mitochondria play a special role in the apoptosis, so they are the major target for the virus related cell death control. The increased permeabilization of mitochondrial membrane and disturbances in the ions concentrations are the early indicators of apoptosis. The analysis of cell cycle may verify the results obtained from the apoptosis-specific tests during EHV-1 neuronal cells infection. For this purpose, the primary cultures of murine neurons established as described before [2] were infected with either Jan-E, Rac-H or 26 strains of EHV-1 (MOI 1.0), and incubated for 24 hours at 37C with 5% CO2.Cells were stained with JC-1 (for mitochondrial transmembrane potential), with VB-48, AO, PI (for changes in the intracellular level of free thiols, accompanying apoptosis or cell damage), and with DAPI (for DNA content quantification, to determine G1/G0, subG1, S and G2/M cell cycle phases), according to manufacturer’s instructions. The results were analyzed using the NucleoView NC-3000 software (details about/concerningthe NucleoCounter NC-3000 design and its capabilities are available at www.chemometec.com). In conclusion, our results have demonstrated that the NucleoCounter NC-3000 can be used in virological studies and the main advantage was its ability to handle a large number of samples with a high degree of precision in viable cells quantitation.

Joanna Cymerys
Warsaw University of Life Science, Poland
Title: Primary murine neurons as in vitro model for investigation neurodegeneration processes caused by human herpesvirus type 2 (HHV-2) infection

Biography: Joanna Cymerys, PhD is currently working as associate professor at Warsaw University of Life Sciences in the Faculty of Veterinary Medicine, Division of Microbiology. She is interested in the field of viral neuroinfections and neurodegeneration.

Abstract: Human herpesviruses type 1 and 2 (HHV-1, -2) are ubiquitous, neurotropic pathogens. They are the most common pathogenic cause of sporadic acute encephalitis in humans. Human herpesvirus encephalitis is associated with a high mortality rate and significant neurological, neuropsychological, and neurobehavioral sequelae, which afflict patients for life. HHV-1 has been suggested as an environmental risk factor for neurodegenerative disease (e.g Alzheimer’s disease). HHV-2 is a closely related virus and it would not be surprising to discover that HHV-2 infection has effects similar to HHV-1 infection on processes connected with neurodegeneration. However, the mechanisms involved in HHV-2 infection that may trigger the neurodegenerative process are still not fully elucidated Most of the available information about the latency establishment, maintenance, and reactivation of α-herpesviruses is derived from in vivo studies, however it is difficult to differentiate specific effects of direct virus-neuron relationship from indirect consequences mediated by immune or non-neuronal supportive cells. In the present study, we have been investigating the potential of primary cultures of murine neurons as an in vitro model for studying the HHV-2 infection. We have been also aiming to answer several key questions regarding cells viability and mitochondrial potential in cultured neurons infected with HHV-2. We have demonstrated that HHV-2 was able to replicate in cultured murine neurons without the need for adaptation and the quantitative analysis of viral DNA (real-time PCR) has shown a high level of HHV-2 replication in neurons. The highest levels of viral DNA in cells were found at 168 hr post infection. Additionally, we have shown a significant increase in the amount of viral DNA in the culture medium at 168 hr post infection, indicating that during long-term HHV-2 infection a continuous production of progeny virions occurs. It has been found that HHV-2 reduced viability of cultured murine neurons and during the infection we have observed a decrease of mitochondrial potential in neurons, especially in the early period of infection. We have also shown that HHV-2 infection of primary murine neurons caused mitochondrial fusion and apparent fragmentation of the mitochondrial network. Moreover, we have noted migration of mitochondria to perinuclear region what might suggest, that they are used by HHV-2 during replication in murine neurons. It might be one of the reason for the development of neurodegenerative disorders.

Jose Gaby Tshikuka
1Department of Family Medicine and Public Health, Faculty of Medicine, University of Botswana, Gaborone, Botswana
Title: Combination antiretroviral therapy regimens and diabetes-relatedcomorbidities among patients attending HIV clinics: a 12-year retrospective cohort study

Biography: Jose Gaby Tshikuka is Department of Family Medicine and Public Health, Faculty of Medicine, University of Botswana, Gaborone, Botswana. He is Doctor of Philosophy: Epidemiology of Infectious Diseases (concentration: Epidemiology, Parasitology, Biostatistics and Nutrition), McGill University, Canada 1996 and also Medical Officer, NIDs Coordinator, Expended Program on Immunization (EPI), World Health Organization.

Abstract: Background:Exposure to combination antiretroviral therapy (cART) is associated with diabetes-related comorbidities (DRC). Botswana has been administering free cART for a long time;however,cART regimens that confer the longest survival to recipientsor that are most associated with DRC, plus biomedical and demographic factors affecting DRC occurrence among Batswana are still ill understood. We aimed to examinecARTassociations with DRC among patients attending HIV clinics in Gaborone, to identify patients’ underlyingbiomedical and demographic risk factors of DRCand, investigate survival of patients on different cART regimens. Methods: Data from HIV clinics were reviewed. Associations between different cARTregimens and DRC were investigated among 531 recipients. Recipients’ DRC risk factors were identified. Cox regression model was run. Unadjusted and adjusted hazard ratios were computed, and hazard and survival functions for different cART regimens plotted. Results: HIV patients on second- and third-line cART were less likely to develop DRC than those on first-line cART;patients with CD4 count ≤ 200 cells/mm3 upon cART initiation were more likely to develop DRC than those who had CD4 count > 200 cells/mm3. Participants who reported adherence to cART were more at risk of developingDRCcompared to those who did not.Overweight patients at initiation to the program had higher risk ofDRC compared with those with normal BMI. Males had lower incidence of the outcome than females and recipients younger than 35 years were less at risk of DRC than those aged 35 years or older. Different cART regimens yielded different survival functions. The shortest survival was among patients on the first-line cART whereas the longest was among patients on the second-line or third-line cART regimen. Conclusion: DRC occurrence among cART recipients is not only a function of the type of cART regimen or the duration of exposure to these drugs but also a function of patients’ underlying DRC risk factors. A review of current cART regimens and a meticulous assignment of patients to specific treatmentregimens will likely improvepatients’ survival. Keywords: HIV patients, combination antiretroviral therapy, underlying risk factors, diabetes-related comorbidities,survival function

Anna Golke
Warsaw University of Life Sciences Poland
Title: Equine PBMCs display sex-specific differences in TLRs gene expression after infection with equine herpesvirus type 1

Biography: Anna Golke is an associate Professor in the Department of Preclinical Science, Faculty of Veterinary Medicine, Warsaw University of Life Sciences. Her scientific interests include neuroinfections and the innate immune response to viral infections, in particular, the use of PRRs ligands in therapy of infectious diseases.

Abstract: Equine herpesvirus type 1 (EHV-1) is a prevalent causative agent of respiratory disorders, abortion and myeloencephalopathy (EHM) in horses. After primary replication in the respiratory epithelium, EHV-1 disseminates through the body via a peripheral blood mononuclear cell (PBMC)-associated viremia. EHV-1 strains with a single nucleotide polymorphism in the viral DNA polymerase gene are statistically more often isolated from cases of EHM and are called neuropathogenic. Equine PBMCs are frequently used as an in vitro model for investigating replication kinetics of neuropathogenic and non-neuropathogenic strains of EHV-1. However, animal sex may influence PMBCs potential to recognize and respond to viral infection. Therefore, the aim of this study was to evaluate mRNA gene expression of two innate immunity receptors: TLR2 and TLR3 in equine PBMCs isolated from mares and stallions and infected in vitro with two different strains of EHV-1. In the present study we have observed differences in TLR2 and TLR3 mRNA genes expression dependent on the EHV-1 strain used for infection as well as the fact, whether PBMCs were collected from mares or stallions. In general, non-neuropathogenic EHV-1 strain showed a capability to stimulate mRNA expression of both investigated genes, when neuropathogenic strain was decreasing it in comparison to uninfected control. Decreased expression of innate immune receptors may impede EHV-1 recognition and support viral replication, what may contribute to the occurrence of EHM. However, we have also observed a significant differences in both TLR2 and TLR3 mRNA genes expression in PMBCs isolated from mares and stallions. It suggest, that this parameter should be taken into account in studies concerning innate immunity responses after viral infection also in research conducted in vitro.

Md. Aminul Islam Apu
Islamic University, Bangladesh.
Title: Zika virus: a threat for future

Biography: Md. Aminul Islam Apu is from Department of Biotechnology and Genetic Engineering, IslamicUniversity, Bangladesh.

Abstract: Zika virus was from on several occasions from amosquitoes named as Aedesafricanus after its discovery in 1947 in Uganda. Zika virus (ZIKV) is an arbovirus belonging to the Flavivirusgenus which was first isolated from a rhesus monkey in a forest of Uganda.Zika virus is the virus which is highly related with some virus like the West Nile viruses. Yellow fever, Japanese encephalitis, and dengueZika virus as it continues its spread throughout the several islands of the West Pacific, non-endemic countries as Japan, Germany, Canada, Australia and United States and in many tropical regions around the world.Zika virus disease, in most causes no or only mild symptoms. While there is no specific treatment for the disease. Sometimesparacetamolcan be effective for the disease. It is one of the major concern that this disease cannot be prevented by any of medications or any of vaccine. Recently the most of the people suffer from a specific problem due to this virus is this disease can spread the pregnant woman’s fetus which gradually causes several birth defects and other brain malformations.This review describes simply about the current understanding the epidemiology,clinical characteristics, transmission, and the diagnosis of Zika virus infection, as wellas the future prospects regarding this Zika virus.

Institute of Medical Virology,School of Medicine of WuhanUniversity, Wuhan China.
Title: Research and development of the Anti Herpes Simplex Virus Effect of Total Flavonoids of Polygonumperfoliatum L

Biography: Mengxin Shen is aPh.D candidate in WuhanUniversity. Her major is pathogen biology and she is mainly engaged in the mechanism of antiviral drug research. Her professor is Zhanqiu Yang whois the director of Institute of Medical Virology, Wuhan University,China.

Abstract: Infectious diseases threat human’s health worldwide, especially for viral diseases. Among those viruses, Herpes simplex virus (HSV) is most ubiquitous and contagious. As a consequence, to develop specific drugs against HSV is a hotspot in the biomedical research. Polygonumperfoliatum L. is named as Gangbangui which had been used as a fork medicine of herpes zoster and inflammation-related therapy. In this study, we try to comprehensively investigate the anti-HSV fractions and mechanisms of Total Flavonoids of Polygonumperfoliatum L. (TFPPL) to provide the solid evidence for clinical treatment of HSV infection. In vitro antiviral tests indicated that TFPPL could directivity inactive viruses and prevent the viral proliferation, but no functions on the viral attachment or penetration. We orally inoculated the 120mg/kg /d TFPPL into Balb/c for 14 days, no obvious toxic symptoms, and no death as well were observed in the animals after 20 days, showing a high safety of TFPPL. In the mice model of HSV infection, we conducted three groups by three TFPPL dose (30mg/kg/d, 15mg/kg/d, 7.5mg/kg/d). Compared with the positive control (acyclovir,ACV, 50 mg/kg/d ) to study the treatment effects of TFPPL in the HSV-1 encephalitis model. There was no difference in statistic with the high dose group, and the high dose group could even prolong the more life time than ACV group .Compared with viral infection group, the number of survival mice and survival time and had a significant difference and in a dose-effect relationship. Therefore,TFPPL could serve as a new candidate for HSV infections.

Oluwakayode Temitope Adekunle
Federal University of Agriculture, Abeokuta, Nigeria.
Title: Detection of enteric viruses and index indicators for evaluating sachet water quality

Biography: Adekunle has completed his MSc in the field of Medical Microbiology and Public Health from the Federal University of Agriculture, Abeokuta, Nigeria and undergraduate studies from Ebonyi State University from the Department of Medical Laboratory Science. He is a Medical Laboratory Scientist at the Obafemi Awolowo University, Nigeria. He has published more than 6 papers in reputed journals

Abstract: Microbiological safety of sachet water remains a public health problem in Nigeria as most brands on quality assessment do not conform to drinking water standards. There is a need to assess sachet drinking water for possible enteric viruses and some index indicators. A total of sixty samples of sachet water were obtained from 5 different brands between February (peak of dry season) and April (rainy season) 2013. Water samples of the same brands were pooled together and concentrated with Polyethylene glycol (PEG) 6000. Viral detection was conducted using Polymerase Chain Reaction techniques targeting specific genes of adenovirus, rotavirus and norovirus. Cryptosporodium parvum, Giardia lamblia and Escherichia coli as index indicators along with other organisms were detected using standard methods. Viral analyses revealed that only one sample which was collected at the peak of dry season tested positive for adenovirus while rotavirus and norovirus were absent in all samples. Adenovirus had a prevalence rate of 20% (1/5) in February and 6.7% (1/15) over three months of collection. No oocyst of Cryptosporodium parvum or ova of Giardia lamblia or any form of parasite was found in all batches of water collected. However, Salmonella enterica serovar Typhi, Pseudomonas aeruginosa, Shigella dysenteriae, and Escherichia coli were detected. Adenovirus was detected by PCR in a sachet water sample that tested negative for Escherichia coli, oocyst of Cryptosporodium parvum and ova of Giardia lamblia. There is the need to screen sachet water periodically for enteric viruses even when they meet bacteriological standards.

Amna Nasir
Department of Pathology & Laboratory Medicine, Pakistan
Title: Identification of Mosquito vectors of Arboviral infections from Karachi, Pakistan

Biography: Amna Nasir is from Department of Pathology & Laboratory Medicine.

Abstract: Background: There is not much data on arthropod-borne viral illnesses in Pakistan. A cross-sectional study has been planned in at different sites in Sindh province of Pakistan aimed to determine the role of arboviruses as a cause of undifferentiated febrile illness in the region. There are two aspects of the study, one to detect Dengue, West Nile, Chikungunya and Japanese Encephalitis viruses in such patients. The other would detect and quantify the mosquito genera in the highly populous city of Karachi and detect the presence of the above-mentioned viruses in the mosquitoes caught from various locations of the city, from May to November 2015. These are the preliminary results of the study on mosquito vectors. Methodology: Mosquitoes were collected from three different regions of Karachi using BG-Sentinel (Biogents®) mosquito trapper. The trapper was set for 12 hours, from 7 p.m. in the evening to 7 am in the morning in Gulshan Town (GT) for 7 days during May 2015, then Saddar Town (ST) and North Nazimabad Town (NNT) for 4 days each during June 2015. Mosquito genera were identified on the basis of antenna, proboscis, palpi and rear end morphology. The insects were then stored in eppendorfs labelled with the assigned number and collection date and frozen at -80oC. The insects will be retrieved later for performing DEN, CHIK, JEV and WNV PCR on their crushed extracts. Results: A total of 54 mosquitoes were caught from the three collection sites, of which 94.5% were culex, 5.6% anopheles and no aedes. In GT the traps, set near potted plants on first floor, yielded the highest number of mosquitoes (6.15/day); 95% of which were culex with 50% females. In ST and NNT the traps were set on second and third floors in balcony and near windows. The mosquito yield was lowest in NNT with only one male culex caught in 4 days, while 2.5 mosquitoes/day from ST. Results of PCR will be available by the end of the study. Conclusion: The predominant mosquito species in Karachi is culex, found primarily near ground level and foliage. This implies easy availability of vector for WNV and JEV. It remains to be seen whether either of these viruses can be detected from mosquitoes trapped from various areas of Karachi.

Dr.Aroor Bhagyalaxmi
Community Medicine Department, B.J.Medical College, New civil Hospital Campus, Asarwa, Ahmedabad , Gujarat India 380016

Biography: Dr.Aroor Bhagyalaxmi is from Community Medicine Department, B.J.Medical College,

Abstract: Abstract: In the post –pandemic period, India has experienced outbreaks of Influenza A(H1N1) in the year 2010, 2011, 2012 and between January to March 2103.This study summarizes the clinical and epidemiological characteristics of all patients with suspected influenza like illness that were hospitalized at a tertiary care center in Ahmedabad, Gujarat from 1st January to 30th April 2103. Total 254 patients with influenza like illness (ILI) were admitted and tested for H1N1 during this period. Data was collected retrospectively about demographic, clinico-epidemiological characteristics, treatment details and final outcomes. Comparison was done between Positive and Negative for H1N1. Out of 254 with ILI, 163 tested positive for H1N1 and 91 were negative. Nearly 50% of the patients were in the age group of 15-45 years in both the groups. Proportion of the elderly patients (>60yrs) were more among positive cases (p=0.0074). Significantly higher proportion of females was reported among positive cases (p=0.0016). However, there was no gender difference in the outcome of both positive and negative cases. There was no significant difference in the fatalities between both the groups (p=0.983)as 41/163(25.15%) positive and 23/91(25.27%) negative patients died. Among H1N1patients, significantly higher mortality was observed among referred patients. Study did not find any significant association between co- morbid conditions and poor outcome in both the groups. Study observed higher deaths among referred cases which could be due to delay in approaching health care facilities, lack of involvement of private sector in early diagnosis and management and also delay in referral services.

Dr. ATHEER aldoori
University of baghdad College of veterinary Medicine Department of microbiology Iraq. Baghdad
Title: The effect of Mangifera extract on different type of tissue culture cells and their antiviral effect on rotavirus .

Biography: Dr.Atheer Aldoori is from University of baghdad,College of veterinary Medicine, Department of microbiology in Iraq Baghdad


DR. Eman Sh. AL-Obeidy.Ph.D
Virology department- PCR unite , Medical city-Baghdad-Iraq.
Title: Prevalence Of Hepatitis G Virus (HGV) Infection And Risk Factors Among Haemodialysis Patients

Biography: DR. Eman Sh. AL-Obeidy.Ph.D is from Virology department- PCR unite , Medical city-Baghdad-Iraq.

Abstract: Background: The hepatitis G virus (HGV),also known GB virus C (GBV-C), is a member of the Flaviviridae family distantly related to hepatitis C virus (HCV) , they infect humans, but is not known to cause human disease. This virus can be transmitted efficiently by blood transfusion and by other parenteral mechanisms and transient and long lasting infections with HGV have been documented in man. Objectives: This study was established to shed light on the prevalence of HGV and investigate the risk factors for viral transmission among haemodialysis patients. Patients and methods: Anti-HCV Ab and Anti-HGV Ab were detected by Enzyme-Linked Immunosorbent Assay (ELISA).HCV RNA and HGV RNA on the other hand, has been detected using PCR technique in the serum of 62 Iraqi haemodialysis patients in comparison with 50 healthy individuals control. Results: Among 150 haemodialysis patients the prevalence of HGV infection was detected in 41.3% in hemodialysis patients by ELISA technique, from those HGV-RNA were observed in 38 (25%) patients. In addition, It was found that blood transfusion increased the risk of HGV infection significantly (P<0.05). However, There was no association between HGV infection and duration of hemodialysis patients. Conclusions: The prevalence of HGV infection in haemodialysis patients seems to be relatively high in our area. Infection with HGV does not seem to play a significant pathogenic role and. only the number of transfusion of blood or blood products increases the risk of new infection during hemodialysis.

Hussam Al Soub,
Communicable Diseases Center, Doha-Qatar
Title: Epidemiology and the changing face of HIV infection in Qatar

Biography: Hussam Al Soub is from Hamad Medical Corporation, Department of Medicine, Communicable Diseases Center, PO Box: 3050, Doha-Qatar

Abstract: Background: To study the demographics, modes of transmission, clinical features and outcome of HIV infection in Qatar and the change in epidemiology over time in the periods before and after 2000. Method: Review of the records of all cases of HIV infection diagnosed in Qatar in the period between 1984 and October 2014. Results: During the study period 306 cases of HIV infection were diagnosed in Qatar. Files were available for review for only 148 patients. Males were more than females with a ratio of 2.5:1. The male to female ratio in those diagnosed after 2000 (group II) was significantly more than in those diagnosed before 2000 (group I). Almost half of the cases were Qatari. The most common mode of transmission was sexual (72%), however in a significant proportion (43%) of those in group I, the mode of transmission was blood transfusion. 54% of patients had a late presentation with an AIDS defining condition or with CD+4 less than 350 cells/ mm3. The mean CD+4 cell count at presentation was 359cells/ mm3, and there was no significant difference between the two groups. Evidence of past infection with HBV, HCV, syphilis and toxoplasmosis was low (10%, 4%, 2.5% and 11% respectively), however the difference between the two groups was significant only for HCV infection. 104 patients (70%) are still alive despite some of them were diagnosed early in HIV epidemic. Conclusion: Qatar remains a low prevalence country for HIV infection. The diseases affects mainly young male adults with many of them presenting late in the disease. The epidemiology of HIV infection in Qatar has changed over time with infection being mostly sexually transmitted in later years. More Non-Qatari are being diagnosed compared with earlier years and this reflects the change in population that occurred in Qatar in recent years. More effort are needed to educate the public especially the young in prevention measures and to improve early diagnosis.

Iman A.El Aziz Khaled
Theodor Bilharz Research Institute (TBRI).Cairo, Egypt
Title: Prevalence of HBV Genotypes in Egypt among Hepatitis Patients

Biography: Iman A.El Aziz Khaled is from Haematology &Blood Bank, Tropical Medicine, Theodor Bilharz Research Institute (TBRI).Cairo, Egypt

Abstract: Introduction It is estimated that 350 million individuals are chronically infected with hepatitis B virus (HBV) and that more than 1 million die from cirrhosis and hepatocellular carcinoma (HCC) each year. HBV has been classified into eight genotypes (A-H) based on the sequence divergence of > 8% in the entire genome, which consists of about 3200 base pairs .Different HBV genotypes have distinct geographical distributions. Africa is one of the highly endemic regions of HBV with five genotypes (A-E) identified: genotype A in Kenya ,genotype D in Tunisia , genotype (A-D) in South Africa and genotype E in Nigeria . According to Egyptian studies , the prevalence of HBsAg in Egypt is of intermediate endemicity (2–8%). Nearly 2-3 million Egyptians are chronic carriers of HBV. Structural and functional differences between genotypes can influence the severity, course and likelihood of complications and hepatitis Be antigen (HBeAg) seroconversion. In addition, HBV genotypes may be associated with differences in response to antiviral therapy. Some studies indicate that HBV genotypes respond differently to interferon in patients with chronic hepatitis B . Aim of the study Our aim in this study was to use the line probe assay (INNO-LiPA HBV Genotyping assay; Innogenetics N.V., Ghent,Belgium) to detect the most prevalent genotypes of HBV among Egyptian hepatitis patients. 2. Patients and Methods The study was approved by the ethical committee of Theodor Bilharz Research Institute (TBRI) (No 52) and informed consents were obtained from patients participating in this study. A total of 140 patients with hepatitis B surface antigen (HBsAg) positivity were enrolled in this study. Of the 140 patients, only 100 patients were HBV DNA positive and those were included in our study and classified into: 20 patients with active hepatitis (AH) diagnosed by HBsAg and HBc-IgM, 75 patients with chronic active hepatitis (CAH) characterized by presence of HBsAg with increased (ALT) level for more than 6 months, and 5 patients with (HCC) diagnosed by ultrasonography. HBV was diagnosed depending on clinical data, liver function tests done by (Hitachi 902), HBV serum markers done by ELISA technique (Abbott AxSYM® HBsAg Assay) and HBV DNA by real time PCR (Two step RT-PCR using Applied Biosystem). Patients were excluded if they were co-infected with hepatitis C virus (HCV) or human immunodeficiency virus (HIV). Results A total of 100 patients with a mean age of 37.17 ± 11.75 years, including 25% females and 75% males, were enrolled in this HBV genotype study. Genotype detection by hybridization of the PCR products to the kit membrane strips was performed as described above. Distribution of HBV genotypes This study showed that HBV infections in hepatitis patients are attributed predominantly to viral genotype D constituted 87% of the total infections. In addition, there was a relatively high prevalence of mixed infections (D/F) represented 13% among the studied group. The Association between liver disease and the prevalence of HBV genotypes was as following: Genotype D was found significantly more often in patients with CAH and HCC than in patients with AH [75/75(100%), (5/5(100%) v (7/20 (35%)]. Mixed infection (D/F) was only found in AH group [13/20 (65%)] (Figure 3). Figure 1. Bands representing oligonucleotide probes specific for HBV genotype in the studied group. Figure 2. HBV genotype distribution in the studied group. Figure 3. The Association between liver disease and the prevalence of HBV genotypes. Conclusion In this study, genotype D was reported as the predominant HBV genotype (87%) in Egypt followed by mixed genotype (D/F) that constituted 13%. Emerging evidence suggests that patients with genotype D infection may develop fulminant hepatitis with high frequency.The existence of HBV genotype F in acute forms of liver disease suggests an association of genotype F with more severe and acute forms of liver disease. Mixed infection was accompanied by acute exacerbation of the chronic disease and may be provoked by population migration. We therefore suggest that HBV genotyping become a routine exercise in clinical medicine and molecular epidemiology.

Jie Wu
Guangdong Provincial Center for Disease Control and Prevention, Guangzhou, China
Title: Effect of Live-Poultry Market Interventions on Influenza A(H7N9) Virus, Guangdong, China

Biography: Jie Wu is from Guangdong Provincial Center for Disease Control and Prevention, Guangzhou, China

Abstract: Since March 2013, three waves of human infection of avian influenza AH7N9 virus have been detected in China. To investigate virus transmission within and across epidemic waves, we used surveillance data and whole-genome analysis of viruses sampled in Guangdong during 2013–2015. We observed a geographic shift of human A(H7N9) infections from the second to the third waves. Live-poultry market interventions were undertaken in epicenter cities; however, spatial phylogenetic analysis indicated that the third wave outbreaks in central Guangdong most likely resulted from local virus persistence rather than introduction from elsewhere. Although the number of clinical cases in humans declined by 35% from the second to the third waves, the genetic diversity of third-wave viruses in Guangdong increased. Our results highlight the epidemic risk to a region reporting comparatively few A(H7N9) cases. Moreover, our results suggest that live-poultry market interventions cannot completely halt A(H7N9) virus persistence and dissemination.

Juliann Nzembi Makau1
Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Japan
Title: Identification of Small Molecule Inhibitors for Influenza A virus Using In Silico and In Vitro Approaches

Biography: Juliann NzembiMakau is a final year PhD candidate in the Program for Nurturing Global Leaders in Tropical and Emerging Communicable Diseases at Nagasaki University, Japan. Juliann received her undergraduate in Medical Laboratory Sciences at Kenyatta University, Kenya and master’s degree in Pharmaceutical Sciences at Nagasaki University. She recently completed three months internship at Texas A&M University, USA. Her research interest is to identify novel targets and antiviral drugs for emerging viral diseases. Her 6 years research work has focused on natural products screening for antiviral activities and structure-based drug design for influenza virus.

Abstract: Influenza viruses are important pathogens that threaten human and animal health. The emergence of resistance to currently available drugs necessitates the development of new drugs with novel mechanisms of action. The virus nucleoprotein is an essential component of the ribonucleoprotein complex for transcription and replication of the virus. In this study [1], we used structure-based approach to find lead inhibitors targeting influenza A virus nucleoprotein. Our work employed an original docking algorithm termed Nagasaki University Docking Engine (NUDE) that ran on the Destination for GPU Intensive Machine supercomputer[2] to select compounds that bind to nucleoprotein from a custom chemical library. Selected compounds were evaluated for antiviral activity in a cell based assay. We found a compound designated NUD-1 belonging to 4-hydroxyquinolinone family that effectively inhibited the replication of various influenza A virus strains including a clinical isolate of 2009 H1N1 pandemic. Analysis of binding between nucleoprotein and NUD-1 using surface plasmon resonance assay and fragment molecular orbital calculations revealed that the compound could bind to nucleoprotein and inhibit protein-protein interactions essential for virus replication. Collectively, our data demonstrate that NUD-1 is a potential lead compound for anti-influenza drug development and highlights the applicability of NUDE in fast selection of important lead compounds. References 1. Makau JN, et al. Identification of small molecules inhibitors for influenza A virus using in silico and in vitro approaches. PLoS One 12(3):e0173582, 2017. 2. Ishibashi D, et al. Structure-based drug discovery for prion disease using a novel binding simulation. EBioMedicine. 9:238–249, 2016.

Kanga Fouamno Henri Lucien
University of Bamenda, Cameroon
Title: The kidney function trends in human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) patients at the Nylon District Hospital, Douala, Cameroon

Biography: Kanga Fouamno Henri Lucien is from Department of Medical Laboratory Science, University of Bamenda, Cameroon

Abstract: Human immunodeficiency virus (HIV) related kidney disease is one of the leading causes of death and affects predominantly people of black descent. Data is unavailable on the presence of kidney disease amongst HIV positive patients in Cameroon and a high prevalence of the disease depicts a high incidence of HIV associated nephropathy. A cohort study was carried out from May to August 2010 at the Nylon District Hospital Douala to investigate the Kidney function trends amongst seropositive individuals. Kidney function tests like serum urea, serum creatinine, creatinine clearance, proteinuria and urine chemistry was measured amongst 329 participants amongst whom 100 (30.4%) were HIV negative and 229 (69.6%) were HIV positive. The age range of the study population was 18 to 60 years, with mean age of 35.122 ± 0.543. There were 94(28.6%) males and 235 (71.5%) females. The percentage of HIV seropositivity was higher in females than in males (74.3% vs. 25.7% p < 0.05). Although, Serum creatinine, creatinine clearance and proteinuria were significantly higher in the control group than in the HIV infected subjects (p < 0.0001, p = 0.046 and p = 0.001, respectively), these values were not indicative of renal pathology. Considering only HIV positive individuals the mean serum creatinine was significantly higher in the Antiretoviral treatment (ART) naïve group when compared to those who were already on ART. These findings indicate that renal function is not affected by the seropositivity status of individuals.

Mazyar Ziyaeyan
Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
Title: Genital tract infections with human papilloma virus, herpes simplex virus, Neisseria gonorrhoeae, and Chlamydia trachomatis in HIV-infected women, Shiraz, Southern Iran

Biography: Mazyar Ziyaeyan is an associate Professor in the Clinical Microbiology Research Center at the Shiraz University of Medical Sciences, Shiraz, Iran where he has been a faculty member since 2002. Mazyar completed his Ph.D. at TarbiatModares University, Tehran, Iran in the field of Medical Virology. He also currently serves as supervisor of viral research/diagnostic laboratory of the research center. During 2014-2015 he was a visiting scholar at Department of Medical Microbiology, Manitoba Centre for Proteomics and Systems Biology, University of Manitoba, Winnipeg, MB, Canada. His research interests lie in the area of rapid diagnostic of viral infection and also evaluation of natural, chemical and biochemical compounds activity on replication of viruses.

Abstract: Objectives. The aim of this study wasto determine the prevalence of human papilloma virus (HPV), herpes simplex virus (HSV), Chlamydia trachomatis (CT), and Neisseria gonorrhoeae (NG), and associated risk factors among HIV-infected women in Shiraz, Iran. Materials and Methods: In this cross-sectional study, from 71 HIV-infected women, aged 17-45 years (mean ± standard deviation: 31.11 ± 6.58 years), cervical swab samples were collected, and tested for HPV, HSV, CT, and NG using PCR assays. Results: Overall, 77.5% were positive for STIs: 36 (50.7%) HPV, 7 (9.9%) HSV, 4 (5.6%) NG, and 27 (38%) CT. In addition, 39 (55%) had only one infection, and 16 (22.5%) suffered from multiple infections. The prevalence of all the respective STIs increased by age, except for HSV which showed a slight decrease. Having low educational level, multiple sex partnership, and be a sex worker were significantly higher among STIs than non-STIs patients. Conclusion: High prevalence of the respective STIs was observed among HIV-infected women in this region. Therefore, significant attention should focus on providing a comprehensive sex education, the use of condoms in order to avoid the spread of STIs to others and themselves,and participation in screening programs for CT and NG are recommended for such high-risk groups. Finally, a nationwide HPV vaccination program should be considered. Keywords: Human papilloma virus, herpes simplex virus, Chlamydia trachomatis, Neisseria gonorrhoeae, HIV.

Melvin Leteane
Plant Vaccine Research Laboratory, Gaborone, Botswana
Title: Expression of Lumpy Skin Disease Virus Proteins in Nicotiana benthamiana

Biography: Melvin Leteane is from Department of Biological Sciences, Plant Vaccine Research Laboratory, Gaborone, Botswana

Abstract: Lumpy Skin Disease Virus (LSDv), genus Capripoxvirus; is an economically important viral disease of cattle in sub-Saharan Africa and more recently with an increasing geographic footprint is spreading to the Middle East and Europe. To date, the control of LSD is by expensive vaccination regimen and sequestration of infected animals. The P32 core protein of LSDv is a promising target for the development of a novel vaccine against LSD as it is highly conserved among Capripoxvirus strains. This study aimed to develop an inexpensive novel LSD vaccine based on Tobacco Mosaic Virus (TMV) coat protein (CP) fusions with LSD epitopes. We report here, for the first time, the transient expression of recombinant expression of LSD P32 peptides in Nicotiana benthamiana. Chimeric constructs were expressed using a TMV-based vector, pJL TRBO, achieving the highest expression level up to 30% of total virus yield of 0.8mg/g of fresh leaf tissue. Analysis of the virus extracts showed a covalent association of the LSD epitopes with the TMV coat protein as well as reactivity to LSD positive sera. The study confirms the utility of P32 core protein as an immunogen for LSD and its relevance for the production of P32 based candidate LSDv vaccines.

Ming Wang
Key Laboratory of Animal Epidemiology and Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing, P.R.China
Title: Influenza A viruses replicate productively in mast cells

Biography: Ming Wang is from Key Laboratory of Animal Epidemiology and Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing, P.R.China

Abstract: The influenza A viruses (IAVs) cause acute respiratory infection in both humans and animals. As a member of the initial lines of host defense system, the role of mast cells during IAV infection has been poorly understood. Here, we characterized for the first time that both avian-like (α-2,3-linked) and human-like (α-2,6- linked) sialic acid (SA) receptors were expressed by the mouse mastocytoma cell line (P815) and human mast cell line( HMC-1). The P815 cells did support the productive replication of H1N1 (A/WSN/33), H5N1 (A/chicken/Henan/1/04) and H7N2 (A/chicken/Hebei/2/02) in vitro while the in vivo infection of H5N1 in mast cells was confirmed by the specific staining of nasal mucosa and lung tissue from mice. The HMC-1 cells did support the productive replication of H5N1 but limited replication of H1N1 in vitro. All the three viruses triggered the infected P815 cells to produce pro-inflammatory cytokines and chemokines including IL-6, IFN-γ, TNF-α, CCL-2, CCL-5 and IP-10, but not the antiviral type I interferon. It was further confirmed that TLR3 pathway was involved in P815 cell response to IAV-infection. Our findings highlight the remarkable tropism and infectivity of IAV to P815 cells, indicating that mast cells may be unneglectable player in the development of IAV infection.

Ryeojin Ko
EwhaWomans University, Korea
Title: Glycogen synthase kinase 3b (GSK3b) regulates TLR3-mediated immune response via downregulation of SOCS1.

Biography: Ryeojin Ko is from Department of Life Science, EwhaWomans University, Seoul 03760, Korea

Abstract: Glycogen synthase kinase 3b (GSK3b) has been reported to regulate TLR3-mediated immune response. However, the mechanisms underlying GSK3b regulation of TLR3-mediated antiviral gene expression remain unclear.Suppressor of cytokine signaling (SOCS) family proteins are negative regulators of inflammatory cytokine and interferon production. Here, we report that GSK3b positively regulates TLR3-mediated antiviral response through inhibiting SOCS1 expression.Overexpression of GSK3b significantly reduced SOCS1 protein level while enhanced antiviral gene expression in a dose-dependent manner.Furthermore, GSK3b physically associated with SOCS1 and mediated SOCS1 ubiquitination.These results suggest that GSK3bregulates TLR3-mediated antiviral response via the downregulation of negative regulator SOCS1

Sahar Sultan Essa
Department of Microbiology, Faculty of Medicine, Kuwait University
Title: Flow Cytometric Analysis of Cellular Immune SubsetsAssociated with Persistent Hepatitis C Virus Infection.

Biography: Dr. Sahar Essa completed her Ph.D from Warwick University, UK in 2001. Currently, she is an academic staff member in the Faculty of Medicine, Microbiology Department, Kuwait University. She is interested in the field of Viral Immunology and published her work in reputed journals. She has been serving as an editorial board member of reputed journals. Member of the European Society for Virology and European Society for Clinical Virology.

Abstract: Background: Hepatitis C virus (HCV) infection is a major public health problem with an estimated 3-4 million people infected each year worldwide (Shepard et al., 2005). 20–30% of individuals acutely infected with HCV will spontaneously clear the virus, with the remaining 70–80% developing persistent HCV infection (Dustin and Rice, 2007). The interplay between the virus and host innate and adaptive immune responses determine the outcome of HCV infection (Rauch et al., 2009). The present study aims to determine the level of cellular immune subsets in HCV-infected patients and to compare it to healthy controls. Methods: This was carried out by investigating the immunophenotypes of immune cells in the peripheral blood of 33 HCV-infected patients and 30 healthy controls before treatments. The immunophenotyping of mononuclear cells in the peripheral blood were evaluated by flow cytometry using antibodies specific to CD3+ (mature T cells), CD3+CD8+ (T cytotoxic cells), CD4+CD25+ (regulatory T cells), CD3+CD4+ (T helper cells), CD8+CD26+ (activated T cells), CD3-CD56+CD16+ (natural killer (NK) cells), CD3+CD56+CD16+ (NKT cells), CD19+ (pan B cells), and CD4+CD25+ (regulatory T cells). Results: There were significantly lower mean values for absolute count and percentage of T lymphocytes (CD3+, P<0.007 & P<0.5 respectively), absolute count of T cytotoxic cells (CD3+CD8+, P<0.005), percentages of regulatory T cells (CD4+CD25+, P<0.001), absolute count of NK cells (CD3-CD56+CD16+, P<0.05), absolute count and percentage of NKT cells (CD3+CD56+CD16+, P<0.001 & P<0.002 respectively), and activated T cells (CD8+CD26+, P<0.5) between HCV-infected patients and healthy donors. On the other hand, insignificantly lower mean values for absolute count and percentage of T helper cells (CD3+CD4+), and B cells (CD19+), were identified among HCV-infected patients and healthy donors. Conclusion: Cellular subsets of the immune system play an important role in the pathogenesis, progression, and clearance of HCV. The screening for multiple cellular markers in the present study may help us to understand the immunopathogenesis of the disease. These findings could lead to new possibilities for immune-based interventions with the aim of restoring functional antiviral T cell responses combined with improved viral clearance.

Seval Bilge Dagalp
Ankara University, Faculty of Veterinary Medicine Department of Virology, Ankara, TURKEY
Title: The molecular and antigenic characterization of Turkish Bovine Herpesvirus type 1 (BoHV-1) isolates

Biography: Seval Bilge Dagalp is from Ankara University, Faculty of Veterinary Medicine Department of Virology, Ankara, TURKEY

Abstract: The Bovine Herpesvirus type 1 (BoHV-1) belongs to the family Herpesviridae, subfamily Alphaherpesvirinae, and is one of the most important pathogens of cattle.The virusis worldwide distributed, with the exception of a few European countries in which the infectionwas eradicated, and causes significant economic losses to the cattle industry. BoHV-1 isolates have been classified into subtypes 1 and 2, based on restriction endonuclease analyses (REA) of the viral DNA after virus isolation. However, all subtypes are antigenically similar. BoHV-1.1 and BoHV-1.2a mostly are related to the respiratory syndrome and abortions. BoHV-1.2b is related to genital infections especially IPV-IPB without abortions. In Turkey, prevalence and etiological role of BoHV-1 in certain kinds of clinical features like respiratory tract infection, mastitis, abortion detected in closed dairy herds have been reported previously. However, there is no knowledge about the BoHV-1 subtypes and the characteristics of the detected BoHV-1. The present study reports the molecular and antigenic characterization of eight BoHV-1 field strains/isolates obtained from the different clinical cases of cattle in Turkey between 1992 and 2014. In this research, we have selected the gC region of BoHV-1 to detect and/or confirm the presence of the virus in the suspected materials and/or after the virus isolation in the MDBK cells. We have compared the detected and/or isolated viruses by phylogenetic analysis, RFLP and VNT techniques. In 4 out of 8 positive samples for BoHV-1 CPEs were observed in MDBK cell culture and thus isolation of viruses was performed. According to the results of sequence analysis, four BoHV-1 field viruses were determined as BoHV-1.1, and others as BoHV-1.2. Moreover, using the RFLP, twoBoHV-1 isolates were confirmed as BoHV-1.1 and other two isolates were confirmed BoHV-1.2 subtype. Differences between BoHV-1.1 and BoHV-1.2 isolates were detected in Virus Neutralization Test (VNT) results with isolates and control viruses (BoHV-1 Cooper strain and BoHV-5 Texas 89 strain). The results showedthe necessityof reliably identifiedBoHV field isolates to better understand epidemiology and pathogenesis of the infection. In addition, it would be useful to identify the subtypes that circulate in the country when vaccination preferences are determined.

Supachai Sakkhachornphop
Research Institute for Health Sciences, Chiang Mai University, Chiang Mai 50200, Thailand
Title: Board-Spectrum Antiviral Activity of Ankyrin Repeat Protein on Viral Assembly Against Chimeric NL4-3 Viruses Carrying Gag-Protease Derived from Circulating Strains Among Northern Thai Patients

Biography: Supachai Sakkhachornphop graduated from Chiang Mai University (CMU), Thailand in 1996. He received a MS in Health Sciences at CMU through a grant from The Johns Hopkins University Fogarty AIDS International Training and Research Program (AITRP). In 2004, he trained in HIV subtyping at Henry M. Jackson Foundation, Rockville, MD, USA. In 2012, he received Ph.D in Biomedical Sciences from CMU. While he was a Ph. D. student, he had a chance to do the research at the Scripps Research Institute, San Diego, CA, USA. He is currently a researcher at the Research Institute for Health Sciences (RIHES), CMU, Thailand.

Abstract: Human immunodeficiency virus (HIV) gene therapy has been anticipated as an alternative treatment for acquired immune deficiency syndrome (AIDS). Recently, the ankyrin repeat protein targeting to HIV-1 capsid domain (CA), AnkGAG1D4, has been selected by phage display and demonstrated the negative effect on viral assembly using HIV-1 laboratory strain NL4-3. The aim of this study was to expand the application of AnkGAG1D4 for inhibiting HIV-1 circulating strains. Herein, we generated chimeric NL4-3 viruses carrying patient-derived Gag-Protease from 131 antiretroviral drug-naïve HIV-1 infected individuals in northern Thailand during year 2001-2012. SupT1 stable T cell line expressing AnkGAG1D4 and ankyrin irrelevent control (AnkA32D3) were challenged with these chimeric viruses. Antiviral activity was evaluated by analyzing the number of syncytium formation and the level of extracellular p24 in culture supernatant of day 7 post-infection (pi). Results were determined and classified by K-means clustering method. Interestingly, SupT1/AnkGAG1D4 significantly showed lower number of infected cells comparing with SupT1/AnkA32D3 which was correlated with the level of p24. This result suggested that AnkGAG1D4 can interfere viral assembly of HIV-1 not only laboratory strain but also the active virus in patients with difference sequences of CA domain. In conclusion, the efficient inhibition effect of AnkGAG1D4-expressing cells to Gag-Protease chimeric HIV-1 supported the possibility of ankyrin-based therapy as an alternative therapeutic molecule for HIV gene therapy in the future.

Xiaoqian Zhang
Division of Animal Infectious Disease, State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University,430070 Wuhan, Hubei, China.
Title: The Selection of Cell Line for PEDV Proliferation

Biography: Dr. Xiaoqian Zhang performes her Ph.D. of preventive veterinary medicine work at State Key Laboratory of Agricultural Microbiology of Huazhong Agricultural University (HZAU) of Wuhan, China. She received her bachelor's degree at the Veterinary Medicine College of HZAU in 2011, in 2012 she joined the Veterinary pathology laboratory of HZAU and started on veterinary pathology research as a master, meanwhile, did some research on the pathogens of poultry bacterial disease and the establishment of rapid and available diagnostic methods. Since 2014, as a Ph.D, her research has been focusing on virologic research, virus-host interaction and vaccine development of swine infectious disease.

Abstract: Porcine epidemic diarrhea caused by porcine epidemic diarrhea virus (PEDV), causing high mortality in sucking piglets, resulting in huge economic losses to swine industry worldwide. Although PEDV was long believed to be replicated in the intestinal epithelial cells in vivo, it was usually difficult to propagate in vitro. Most researchers chose Vero cells as the model cell line, while it is deficient in interferon production and not a cell line originated from non-homologous species. Four different cell lines (Vero cells, Marc-145 cells, ST cells and IPEC) were compared to select an appropriate model cell line. The Vero cell line was used for propagation of PEDV strain CH/YNKM-144/2013 (GenBank: KF761675.1). Viral propagation was confirmed by daily observation of the cytopathic effect (CPE), and indirect immunofluorescence assay (IFA) using a monoclonal antibody against PEDV S protein. Meanwhile, the nucleus of these cells were stained with DAPI. All cell lines were transfected with the pEGFP-N1, using lipofectamine™ 2000 reagent. CPE was appeared at 8hpi in all cell lines except IPEC. At 12hpi, the PEDV infection rate of Vero cells, Marc-145 cells and ST cells almost reached at 40%, 18%, 80%, respectively. IPEC never exhibited any visible CPE or the green fluorescence. The pEGFP-N1 transfection into ST cells generated strong green fluorescence signals. At 12hpi, the transfection efficiency can reach 50%. The results demonstrated that ST cell line can also be used as a model cell line or the subsequent interferon-related in-vitro experiments of PEDV.

Xu Yi
Department of Infectious Disease, Guangzhou Women and Children’s Medical Center, Guangzhou 510120, China.
Title: Immune Response and Gut Microorganisms in Children with Differing Severities of Hand, Foot, and Mouth Disease

Biography: Xu Yi is from Department of Infectious Disease, Guangzhou Women and Children’s Medical Center, Guangzhou, China.

Abstract: Hand, foot, and mouth disease (HFMD) is a common childhood infectious disease caused by human enteroviruses. Some patients, particularly young children, may have severe neurological or cardiac complications, causing death in some cases. However, the pathological mechanism leading to severe HFMD is not fully understood, and the severity of HFMD is difficult to predict. In this study, we hypothesized that the disease outcome would be influenced by the patient immunological status which reflects the combination of patient genotype and living environment/condition. RNA sequencing (RNA-Seq) was used to analyze the whole transcriptome of peripheral blood mononuclear cells (PBMCs) isolated from patients with severe and mild disease, as well as healthy age-matched controls. The results showed that the genes involved in immune response-related pathways, especially in the innate immune pathways, were significantly up-regulated in mild cases but were down-regulated in severe cases, indicating that the innate immune response plays an important role in antiviral infection and in controlling progression of the disease. To explore the mechanism of the insufficient immune response in severe cases, we sequenced the human major histocompatibility complex (MHC) region, and identified four expression quantitative trait loci (by eQTL) that were significantly associated with the severity of HFMD. These results suggest a causal model in which an individual’s genotype influences their susceptibility to infection of human enteroviruses through changes in gene expression. Moreover, we analyzed the meta-transcriptome of HFMD patients and found that the differentially expressed genes in the gut were associated with the insufficient immune response in severe cases. Meta-transcript linkage analysis revealed that several gram-negative bacteria such as Escherichia coli and Bacteroides enriched in severe cases, and showed the enrichment of metabolic pathways in severe cases, implying that enrichment of certain bacteria may facilitate viral infectivity. This integrative study combines genetic, transcriptional, and gut meta-transcriptome data to uncover the mechanisms affecting the severity of HFMD. Furthermore, we identified a series of biomarkers based on PBMC RNA-Seq, MHC polymorphism, and gut meta-transcriptome data, respectively, facilitating prediction of the severity of HFMD at the early stage of the disease.

Yasir Arshad
National Institute of Health, Islamabad Pakistan and COMSATS Institute of Information Technology, Islamabad, Pakistan
Title: Title: Molecular Characterization of Human Metapneumovirus (hMPV) Among Children of Less Than 5 Years Age in Islamabad


Abstract: Lower respiratory illness is the leading cause of deaths among children in both low and high income countries. Deaths due to respiratory disorders have been increased in South Asia and sub-saharan Africa. In USA, around 3% children are hospitalized due to severe respiratory illness every year. Human Metapneumovirus (hMPV) is one of the contributors of respiratory illness among children as it causes serious problems which ranges from upper respiratory tract illness to lower respiratory tract illness in the form of bronchiolitis and pneumonia. It causes inflammation and necrosis of epithelial lining of bronchioles and result in sloughing. Two genotypes were frequently reported around the world circulating among populations with two sub-groups each. ELISA and PCR are used frequently to detect the presence of hMPV among children showing symptoms of acute respiratory illness. Real-time PCR was used for the screening of throat swabs taken from suspected samples and two genes have been sequenced which are F and G involved in fusion of virus to host membrane and glycoprotein assists in fusion process. 16.5% suspects were positive for the presence of hMPV of which A2a (10%), A2b (20%), B1 (10%) and B2 (60%) genotypes were found which represent two genotypes A, B, groups and two lineages A2a and A2b. Phylogenetic analysis was performed using Kimura-2 parameter model which shows that the study strains were closely related to the strains reported previously from Pune and Kolkata, India, China and prototype strains from Netherlands. Co-circulation of both genotypes and groups were evident in this study.

Yi-dong Mao
School of Medicine of WuhanUniversity, Wuhan,P. R. China
Title: Establishment of diarrhea model with rotavirus infection in ICR mice

Biography: Yi-dong Mao is 25 years old. She is in pathogenic biology major at Wuhan University Graduate School, studied under professor Zhan-qiuYang , engaged in research on drug - resistant virus.

Abstract: Objective:Infantile diarrhea caused by rotavirus (RV) is the most common illnesses in child. Establishment of diarrhea model is important for the Study of rotavirus infection mechanisms and evaluation on effect of drug treatment. Methods:4d,7d,21d ICR mice were used RV (G3 type 709 strain ) inoculation via oral approach. The control group into virus - free culture medium.Observe the infection situation in daily, respectively, ELISA detection of fecal RV antigen, and pathological examination of the small intestines. Results: 4d and 7d suckling mice diarrhea after 24h inoculation of virus, 21d neonatal mice were not observed symptoms.2-3 days after infected rotavirus, severe diarrhea, watery visible, spirit is dispirited. Stool ELISA test RV antigen was positive, histopathological examination revealed,in experimental mice,lesions of the small intestine in the expanded tip of the villus. Most of the injured part at the top of the villi, myenteronslightly thinner than normal, small intestinal goblet cells decreased, autopsy found that a lot of light yellow liquid manure pile up in the colon, also found that the brittleness of the intestinal tissue is relatively large, fragile.Normal Neonatal mice Small Intestine with Tan formed stool in the colon, intestinal tissue are elastic. Diarrhea in suckling mice can be recovered completely within 7d. Conclusion: successful infection in 4d and 7d ICR mice with rotavirus,provide means for pathogenicity and in vivo Pharmacodynamic studies.

Mohammadi Mashallah, M.B.B.S., M.D.
Iranian Research Institute of Plant Protection, Iran.
Title: A truncated C-terminal fragment of Mycobacterium tuberculosis HSP70 enhances cell-mediated immune response and longevity of the total IgG to influenza A virus M2e protein in mice

Biography: Working Experience: Identifying & collaboration with partners in line with institute activities, Creating and expanding communication and co-operations. Organizing workshops/seminars/ Conferences/ Meetings Preparations of proposals and Annual reports. Arranging bilateral research/ education opportunities. Staying abreast of relevant knowledge in the field. Proactively exploring the sources of bilateral research funding

Abstract: As the importance of virus-specific IgG2a and strong induction of Th1 type immune response for virus clearance was reported, conventional influenza vaccines induce a highly humoral immune response and fail to induce cytotoxic T-lymphocyte (CTL) immunity. Hence, in agreement with heat shock protein 70 (HSP70) acting as Th1 cytokine-like adjuvant, an Escherichia coli-expressed r4M2e.HSP70c fusion pro- tein comprising C-terminus of Mycobacterium tuberculosis HSP70 genetically fused to four tandem repeats of influenza A virus M2e was constructed. Then, the case-control study was carried out to evaluate the humoral and cellular responses elicited against M2e in Balb/C mice by intramuscular immunization with r4M2e.HSP70c alone. Our results showed that r4M2e.HSP70c rather than control groups, r4M2e, r4M2e + Alum, or HSP70c, significantly elevated both longevity and serum level of the total M2e-specific IgG antibody, induced a Th1 skewed humoral and cellular immune responses, increased the level of IFN-c in BALF, and promoted the proliferation of peripheral blood lymphocytes. Furthermore, a virus challenge experiment revealed that mice vaccinated with r4M2e.HSP70c limited the severity of influenza A disease by 100% survival rate, less sever body weight loss and delaying the onset of morbidity in mice for 2 days rather than other control groups. Here, we used r4M2e.HSP70c to stimulate M2e-specific antibody and cellular immune responses in Balb/C mice. The mHSP70c in the fusion form induced a long lasting Th1 skewed humoral and cellular immune responses against its associated protein. It seems anti-M2e antibodies limit viral replication and ameliorate influenza infection that allows the immune system to induce sterilizing HA-antibody against whole virion that leads to full protection against virulent influenza infection.

Liang-jun Chen
Wuhan University, Wuhan,China.
Title: Characterization of the pathogenicity of novel Hantaan virus and the efficacy of its inactivated vaccine on Hantaan virus infection

Biography: Liang-jun Chen is a PHD student from Wuhan University. During the doctoral period, his research was focused on the pathogenicity of novel Hantaan virus and the epidemiology of avian influenza viruses.

Abstract: Hantaan virus (HTNV) is the main etiologic agent responsible for hemorrhagic fever with renal syndrome (HFRS). The occurrence of novel hantaan virus may pose potential danger to the control and prevention of HFRS in China, which highlights the significance of vaccine development in public health management. In the present study, initial biological characterization and pathogenicity of HV 76-118, HV114 and novel isolated HV004 from the epidemic areas of Hubei province was performed in susceptible cells and in vivo. HV004 had similar but higher infectivity and pathogenicity to that of HV76-118 and HV114 in suckling mice. An experimental HV004 inactivated vaccine was later prepared, and its corresponding immunogenicity was analyzed in Balb/C mice. The results showed that vaccination intraperitionally (i.p.) with inactivated HTNV vaccine adjuvanted with aluminum followed by an i.p. challenge with 106 PFU/ml Hantaan virus afforded full protection to Hantaan virus challenge. All the immunized mice of every group elicited serum neutralizing antibody with increasing dosages, which could protect Vero-E6 cells from HTNV infection. A dose-dependent stimulation index of splenocytes could also be observed in immunized mice. These findings suggest that inactivated Hantaan vaccine could stimulate mice to produce high levels of antibodies with neutralization activity. Our data also demonstrated that the percentage of IFN-γ-producing CD3+CD8+ T cells was significantly higher in the spleens of immunized mice than that of control. All these findings directly show that the vaccine could elicit specific anti-Hantaan virus humoral and cellular immune response in Balb/C mice against prevalent strain of Hantaan virus in central-south of China

Virology Congress 2017 | by: Scientific Future Group