Day :
- Sessions: General Virology and Basic Science | Veterinary Virology | Viral Hepatitis | Therapeutic Approaches for Viral Infections | Agriculture and Plant Virology | Viral Pathogenesis,Virulence & Latency | Advances in Drug Discovery
Location: Greenspring 1
Chair
Rob Striker
University of Wisconsin, USA
Co-Chair
Mugdha Vasireddi
Georgia State University, USA
Session Introduction
Rob Striker
University of Wisconsin, USA
Title: Monitoring the latent HIV reservoir and immune recovery in patients on therapy
Time : 12:00-12:20
Biography:
Rob Striker has completed his MD/PhD from Washington University in St. Louis and was an Infectious Disease and Howard Hughes Medical Institute Postdoctoral Fellow at Stanford University School of Medicine. He, now sees patients for the Veterans Association, the University of Wisconsin, and AIDS Resource Center of Wisconsin. He has published more than 45 papers and runs a research lab working to improve how antivirals are developed and prescribed?
Abstract:
One person out of 30 million has had a sterilizing cure of HIV. While this cure came through a risky stem cell transplant, it demonstrates that cure of HIV is possible. There is general agreement that latently infected CD4 memory T cells are now the major barrier to a cure, but no agreement on how to measure this “reservoir”? Furthermore, the compartment that is easiest to serially sample (blood) likely only contains less than 10% of the reservoir, since most of the reservoir is dispersed in different lymph nodes. Our goal is to correlate blood reservoir measurements with a clinical marker (CD4:CD8 ratio) that is already correlated with immune health and risk of death. We have decreased the time to answer and improved the automation of two types of reservoir measurements, TILDA (Tat/rev Induced Limiting Dilution Assay) and Quantitative Viral Outgrowth Assay (QVOA). The TILDA assay, we have improved the signal: noise by adding a patented sample prep step developed by Salus Discovery. We have quantified ~300 CD4:CD8 ratio responses over decades and are testing the hypothesis that there will be a correlation between reservoir size in the peripheral blood and ratio responsiveness. Rigorous analysis will be needed to determine how the easily available CD4:CD8 ratio?, may (or may not) be related to the circulating reservoir, but circumstanial evidence already exists and will be discussed
Shubhada Bopegamage
Slovak Medical University, Slovak Republic
Title: Necessity of modifying the angle of viewing viral infections during pregnancy
Time : 12:20-12:40
Biography:
Shubhada Bopegamage work is focused on the pathogenesis and diagnosis of enteroviruses. She received her BSc Microbiology degree from Pune, India and MSc Microbiology degree from Mumbai, India. She got her PhD in Biological Sciences from the Academy of Medical Sciences, Moscow, Russia. She is known in the Enterovirus research, since 2005 for her work on the in vivo experimental coxsackievirus oral infection of mice, and experimental coxsackievirus infection during gravidity. She is involved in research and teaching and has guided several MSc and PhD students. She has coordinated and has lead several national and international projects as a principal or co-investigator.
Abstract:
Infections, which occur during gravidity, may result in intrauterine transmission of the pathogen causing fetal damage, early pregnancy loss/early miscarriage, and spontaneous or preterm births. The uterine microbiome during pregnancy influences the course of the immune development of the child and the gut microbiome of the neonate. The outcome depends on fetal gestational age at the time of infection, maternal immunological and nutritional status, and other factors: smoking, drugs, stress and alcohol abuse during pregnancy and the infecting agent and its pathogenic potential. Viral infections during pregnancy are often asymptomatic without risk of fetal damage. However, some viruses may interfere with the embryonic development or cause a severe fetal damage and congenital malformations of different types affecting various fetal systems (urological, cardiac) resulting in neonatal diseases with severity ranging from mild to transient symptoms or fatal outcome. Accepted intrauterine vertically transmitted infections are Toxoplasma gondii, Rubella virus, Cytomegalovirus, Herpes simplex virus-2 infections, known by acronym TORCH, where ‘O’ was at first "TO" for toxoplasma. Recently ‘O’ represents ‘other infections’: Enteroviruses, Varicella zoster virus, Chlamydia, HIV, Human T-lymphotropic virus, Treponema pallidum, Zika virus, Hepatitis B, D, and E viruses, and Parvovirus B19. Another suggestion was acronym CHEAPTORCHES which included Chickenpox (Varicella Zoster), Hepatitis B, D viruses, Enteroviruses, AIDS (HIV), Parvovirus B19, where ‘O’ stands for Streptococcus, Listeria, Candida and Lyme disease Neisseria gonorrhoe, Chlamydia, Ureaplasma, Human papillomavirus and Treponema pallidum. We suggest that the original acronym TORCH should be modified in the present days. Additional viruses (where infections may be incidental) and placental role during viral infections and protection of the fetus against pathogens should be considered. The perpetual aspects of poverty, modern dietary/nutritional changes, and emerging and re-emerging viruses call for a reconsideration/modification of the antenatal management of viral infections, and research on the potential mechanisms of infection during pregnancy
Nickolai Nossik
The D.I. Ivanovsky Institute of Virology, Russia
Title: Resistance of DNA and RNA viruses to UV radiation
Time : 12:40-13:00
Biography:
Nickolai Nossik graduated from Moscow Medical University as a Physician. He did his PhD in 1960 and Doctor of Science in 1988. From 1968-1973, he was the WHO expert in SEARO (India) on national and international projects on Virology and Epidemiology. In 1975, he was as a Visiting Scientist and was doing the research study on oncornaviruses at the National Institute of Oncology (Fort Detrick). His main scientific interests are - interferons, immunomodulators, antiviral and virucidal preparations. He is the Head of Laboratory of Virus Ontogenesis Ivanovsky Institute of Virology, N.F. Gamaleya FRCEM», Moscow, Russia. He has published 2 monographies, 6 books chapters and more than 200 papers in scientific journals
Abstract:
The inactivation of different kind of pathogens by UV radiation is known for long time and widely used. Due to wide variety of viruses (by structure, size, envelope) their sensitivity to UV radiation is quite different and not quite predictable. DNA viruses (adenovirus, type 5, Herpes simplex virus (HSV), type 1) and RNA viruses (human immunodeficiency virus (HIV), type 1, poliovirus, type 1, Sabin strain) were obtained from State collection of viruses (the D.I. Ivanovsky Institute of Virology). The source of UV radiation was a 15 watt low pressure mercury vapor lamp (254 nm over 60%). The samples were placed 0.3 m direct under the UV lamp flow. The dose of inactivation D (Jm2) 90.0% - 99.99% was estimated. It was shown that poliovirus and Herpes simplex virus were not very resistant to UV radiation (D99.99 =128.7 Jm2 for poliovirus and 193.0 Jm2 for HSV). Adenovirus was much more resistant and kinetics of inactivation was different (D99.99=772.0 Jm2). HIV was most resistant to UV radiation among the studied viruses. It takes more than 4hrs to inactivate virus onto surface (D99.9 > 1500 J/m2). The results of the study indicate that there is no direct connection between sensitivity to UV light and the size of the virion or presenceabsence of the envelope. The diverse resistance of the different viruses to UV radiation should be taken into account, when UV light is used to inactivate infectious viruses in hospitals and other public environments
Sharad Kumar Yadav
DUVASU, India
Title: Lytic bacteriophages as specific and selective probe for detection of bacterial pathogens
Time : 15:00-15:20
Biography:
Sharad Kumar Yadav has 28 years of teaching and research experience and has served to various senior positions of the University including Registrar of the DUVASU University. He is currently a Professor, Head of Department of Veterinary Microbiology, and the Director at Cow Research Institute at DUVASU, Mathura India. He has published number of papers in reputed International and National journals and has a vast experience in the arena of BHV-I virus
Abstract:
Bacteriophages are proven viruses to be valuable tools in the fight of mankind and animals against diseases and infections. Being host-specific these bacterial viruses are species specific, self-perpetuating, self-limiting and eco-friendly in nature targeting only specific bacteria as per their host range. This property of lytic bacteriophages has been explored in detecting the bacterial pathogens in a new technique in microbiological laboratories for confirming the causative agents as specific bacteria and the technique is known as “Phage typing”. Phage typing has become a popular tool to differentiate bacterial isolates, hence currently used very widely in epidemiological studies to identify and characterize the outbreak-associated strains of Salmonella, Campylobacter, Escherichia, Listeria and many other bacteria based upon receptor specificity. Electron microscopic studies of various lytic phages against bacteria such as Proteus, Bacillus and T-4 bacteriophages; sewage coliphages along with morphotyping of bacteriophages of Lactobacillus helveticus have been already done. Ten varieties of morphologically different phages have been classified into four different families; namely, Myoviridae, Styloviridae, Podoviridae and Microviridae; whereas based on morphology and nucleic acid content additionally Microviridae, Inoviridae, Siphoviridae and Leviviridae are taken into consideration. Bacterial phage typing is an interesting tool directly based upon whole bacterial cells and hence no much expensive instrumentation or skilled man power is required. Due to its practical benefits, this typing technique has gained the importance and popularity in the science of bacteriology
Mugdha Vasireddi
Georgia State University, USA
Title: Impairment of NK cell activation and function during Herpes B virus infection
Time : 15:20-15:40
Biography:
Mugdha Vasireddi has her expertise in studying host-pathogen relationship between humans and their pathogens. Her novel cell culture model represents an in vivo system that could be utilized, where an appropriate animal model cannot be used to study host immune responses. She is also interested in developing rapid and accurate diagnostics for emerging and re-emerging viruses such as influenza and Zika virus. Currently, she serves as one of the Clinical Lab Directors for National B Virus Resource Center that tests human and non-human primate samples for herpes B virus infection. She is part of a team that is focused on developing and extending diagnostic services for Zika virus detection
Abstract:
Herpes B virus, endemic in macaques, is a simplex virus like HSV-1 in its disease manifestation in natural hosts. Zoonotic infection of herpes B virus in humans, however, results in 80% fatality without timely antiviral intervention. Our previous studies revealed that herpes B virus infected human cells do not down-regulate MHC Class I molecules on the surface of infected cells, but rather up-regulate NK cell inhibitory MHC Class Ib molecules, HLA-E and HLA-G. This induction of NK cell inhibitory markers is an indicator of impairment of NK cells. Therefore, we hypothesized that herpes B virus escapes natural killer (NK) cell lysis in zoonotically infected humans by inhibiting NK cell activation. To test this hypothesis, we used a cell culture model system to examine whether herpes B virus infected HFF cells and herpes B virus infected HFFs co-cultured with peripheral blood mononuclear cells (PBMCs) generated the production of NK cell-activating cytokines and chemokines. Analysis of results revealed that herpes B virus infection resulted in down-regulation of NK cell-activating cytokines and chemokines, i.e., IFN-α, IP-10, IL-8, and MCP-1. Next, we measured NK cell activity during herpes B virus infection by quantifying the expression of cytotoxic granules (granzyme and perforin) and cytokine (TNF-α and IFN-γ) expression. Our results demonstrated that NK cells with or without stimulation during herpes B virus infection failed to express cytotoxic granules or cytokines supporting our hypothesis that NK cell activity is impaired during human infection. In addition, we also examined herpes B virus titers in the HFF cells co-cultured with IFN-α stimulated or un-stimulated NK cells. Our results suggested that NK cells were incapable of restricting herpes B virus infection even when stimulated with IFN-α. Therefore, these results suggest that NK cells during B virus infection are rendered inactive
Zenab Aly Torky
Ain Shams University, Egypt
Title: Pharmacological evaluation of the Hibiscus herbal extract against herpes simplex virus-type 1 as an antiviral drug in vitro
Time : 16:00-16:20
Biography:
Dr.Zenab Torky is an Assistant Professor of Microbiology in the department of Microbiology, Faculty of Science, Ain Shams University, in Egypt. She also worked as a Visiting Scientist in the department of Biological Sciences, University of Louisville, USA. Dr.Zenab has also reviewed and edited many papers for the Food Safety Journal.
Abstract:
High prevalence of viral infections and constant emergence of resistant virus strains worldwide calls for an urgent need to discover/develop novel therapeutic agents to improve the global treatment of virus infection especially against Herpes Simplex Virus-type 1 (HSV-1). The objective of this study is to conduct phytochemical screenings to have an insight on the identification, the essential components of the methanolic/aqueous Hibiscus extracts, cell viability and cell proliferation assay as well as the antiviral activity of the methanolic/aqueous of Hibiscus were evaluated by reduction of the viral cytopathic effect, which showed a potential activity of both extracts against HSV-1 as indicated by selectivity index (SI) values of 8.0 and 7.7 for methanolic and aqueous Hibiscus extracts correspondingly. Different experiments like viral binding, viral adsorption, virucidal, and time of addition assays were designed to investigate the mechanism of antiviral activity of Hibiscus extract revealing that the extract not only prevented the virus particles from interacting with the Vero cells in the pre-treatment assay but also had a prophylactic effect blocking the replication of the virus causing prophylactic selectivity indices of 6.1 and 5.2 for methanolic and aqueous Hibiscus extracts correspondingly, confirming a maximum protection of Vero cells against HSV-1 attack. Results shown in this study demonstrate the potent and broad spectrum antiviral activity due to the multiple components contained in the Hibiscus extract from active ingredients. Fluorescence microscopy were used to demonstrate the viral kinetics which revealed that the antiviral activity of all Hibiscus extracts tested was effective during the first stages of the virus replication cycle and thus, Hibiscus extract may provide a novel treatment for HSV-1 infectivity
Filippo de Nicolellis
Friuli Venezia Giulia, Italy
Title: The communication about and the availability towards vaccinal campaigns against influenza in primary health care in north east Italy: outcomes and considerations four years later
Time : 16:20-16:40
Biography:
Filippo de Nicolellis has Graduated at Rome University “La Sapienza” in 1986, where he specialized in infectious diseases in 1990. From 1995 to 2000 he has worked as a family doctor in Doberdò del Lago/Doberdob (Gorizia) and since 2000 in Fiumicello, Udine, in Friuli Venezia Giulia, in the north-east of Italy. Since 2000, he has been the President of the Medical Association “Croce Medica”, which deals with the organization of Health Services and Permanent Training and particularly with Primary Health Care and Emergency
Abstract:
You will be introduced to a four-year project about developing e-communication and other tools in a rural area of Friuli Venezia Giulia, Italy. The Medical Association “CROCE MEDICA” is promoting the use of posters in family doctor’s offices as well as of e-mails to communicate the patients dates and procedures for the vaccination against influenza. Furthermore, the association e-mails, its registered doctors, a dispatch about influenza as well as a request for information about the illness seasonal evolution in the various areas of Friuli Venezia Giulia. Doctors have shown resistance in developing e-communication with their patients; moreover, there are still problems with patients’ acceptance of vaccination due to the spread all around Italy of theory against any forms of vaccine. However, there has been an increase in the percentage of vaccinated people in the investigated area in the last years. I believe all efforts aiming at improving the communication with patients and explaining how to obtain the vaccination against influenza are useful in any case
G M Vennila
Sri Ramachandra University, India
Title: NRDMC-BBTV non-radioactive detection and molecular characterization of Banana bunchy top virus Chennai isolate
Time : 16:40-17:00
Biography:
Ms. Vennila has currently pursuing full time Ph.D in Sri Ramachandra University, Chennai, INDIA and completed her M.sc in Life Sciences and M.Phil in Biotechnology from Bharathidasan University in the year 2005. She worked as a Lecturer for five years before joining Ph.D. She has attended many national and international seminars and conferences. She has won first prize in oral presentation in a national seminar (2010).
Abstract:
Many pests and diseases have significantly affected Musa cultivation. Viral diseases are even more devastating for banana plantation. One of the most serious viral diseases of banana is the Banana Bunchy top disease caused by Banana bunchy top virus, a multicomponent, circular, single stranded DNA virus belongs to the family Nanoviridae. Apart from BBTV, Cauliflower mosaic virus (CMV), Banana streak virus (BSV), Banana Bromo Mosaic virus (BBrMV) also infects banana. Banana is the only host of BBTV and P. nigeronervosa is the only vector that carries BBTV. Detection of BBTV using slot blot hybridization of DNA and RNA with gene specific probes showed variation at DNA level. Comparatively, RNA was better in detecting the viral components than DNA. We have cloned and sequenced all the six full length components of BBTV isolate from Chennai region. Sequence data shows that the Chennai isolate belongs to South-Pacific group of BBTV and have highest sequence identity to isolates from this group rather than Asian group members.
Rafael A Guerrero R
National University of Colombia, Colombia
Title: Evaluation of oncolytic use of Rotavirus Wt1-5 in acute lymphocytic leukemia REH cell line
Time : 17:00-17:20
Biography:
Rafael A Guerrero R is a Doctor in Medicine with a Master’s degree in Biochemistry from the National University of Colombia. He is a current PhD candidate in Biotechnology at the same university. He is also a part time Lecturer of Biochemistry at the Department of Physiological Sciences, School of Medicine, National University of Colombia and at the School of Medicine, Health Science University Foundation. He has published some papers about of Rotaviruses.
Abstract:
Oncolytic viruses are viruses selected to infect and replicate specifically in tumor cells and then decreasing cell viability. Some viruses have been adapted or modified to give them enhanced infectivity and selectivity toward target tumor cells. Oncolytic viruses have become a novel approach for treating some cancers by inducing cell lysis or immune response to tumor cells. In present work, acute lymphocytic leukemia REH cells was infected with adapted Wt1-5 Rotavirus previously reported. Infection produced by active oncolytic Rotavirus induced expression of cell death signals in cancer cells but not in non-tumor cells. Cell death was characterized using DiOC6/7-AAD, DNA fragmentation test, Annexin V, TUNEL and Caspase assay. We propose that the oncolytic properties of this Wt1-5 Rotavirus are derived from the fact that it specifically targets cells expressing heat shock proteins, protein disulfide isomerase and αVβ3 integrin on their cell surface, which is common in many cancer cells. The interaction of Rotavirus and cell surface proteins was evaluated with flow cytometry and confocal microscopy. The results suggest that the Wt1-5 Rotavirus tested in the present work has oncolytic properties that allow us to propose that this adapted Rotavirus could be assayed as an oncolytic virus in other cancer cells, including leukemia and animal models.
- Workshop
Session Introduction
Abdelouaheb Benani
Pasteur Institute of Morocco, Morocco
Title: Hepatitis B and C: Epidemiology, treatment and prevention strategy
Time : 14:00-15:00
Biography:
Abdelouaheb Benani is the Head of Molecular Biology Laboratory at Pasteur Institute of Morocco which is a national reference Molecular Biology Laboratory for Hepatitis C RAMED Program (Moroccan Ministry of Health). He is also the Elect-President of Arab Society for Virology, 2nd Vice-President of Moroccan Society for Virology and an Active Member on the Advisory Board, Consultative Meeting on Regional Strategy for Prevention and Control of Viral Hepatitis and Other Epidemics-Prone Blood-Borne Diseases, in 2008 (WHO/EMRO, Cairo, Egypt). In 2002, he got his PhD in Molecular Microbiology (Fes, Morocco) on the Molecular Epidemiology of Hepatitis C in Morocco. In 1993, he got his Master of Molecular Biology and Biotechnology at Université Libre of Bruxelles (ULB, Belgium) in anti-tumoral action of Parvoviruses. He is an Active Researcher in molecular epidemiology of HBV and HCV in general population and high-risk groups in Morocco. He has organized the 1st International Symposium for Virology in Morocco in 2003 (Marrakech, Morocco), the First PCR Forum and Molecular Typing at Pasteur Institute of Morocco, and the Hepatitis C Workshop at the 5th World Congress on Virology (Atlanta, USA 2015). He is implicated in student’s research for their training, Master and PhD. He has participated in several international and national workshops and congresses. He has published in several international journals including Journal of medical virology, BMC public health, Virology Journal, European Journal of Public Health. He serves as a Reviewer as well as Editorial Board Member of some international journals. Currently, he is a reviewer of some grant agency.
Abstract:
Hepatitis B virus (HBV) and hepatitis C virus (HCV) infections are major public health issues worldwide and the leading cause of chronic liver disease, cirrhosis, and hepatocellular carcinoma. Globally, these infections are responsible for over 1.4 million deaths annually. Approximately, 64-103 million and 240 million people around the globe are chronically infected with HCV and HBV respectively. HCV, which belongs to Flaviviridae family, is highly variable and classified into six major genotypes further divided into more than 67 subtypes. HCV genotyping is correlated with disease severity, therapy response and is a critical factor in determining the optimal treatment duration. Furthermore, considerable progress has been achieved in the efficiency of therapy outstandingly improved by the development of new drugs and new therapeutic strategies. The availability of direct acting antivirals (DAA) treatments has tremendously improved HCV cure rates, which now exceed 95% in most patient populations. HCV epidemic in north Africa falls into an intermediate range, characterized by a decreasing east-west gradient, with the average seroprevalence estimated between 1.2 and 1.9%. HBV epidemic is more rampant, with countries classified as highly or intermediately infected. Egypt harbors, the highest HCV prevalence globally, hovering around 14.7%, with 15% of the total population having been infected, since the parenteral antischistosomal therapy implemented in the 1920s and an HBV prevalence averaging 4%. Libya borders Egypt, yet has a lower endemicity for both HBV and HCV, estimated at 2.2 and 1.3%, respectively. Tunisia and Morocco are intermediate for both HBV and HCV, with rates for HBsAg falling between 4-7% (Tunisia) and 1.81% (Morocco), and anti-HCV seropositivity at 0.4 (Tunisia) and 1.58% (Morocco) for the general population and 60% was found among intravenous drug users. The majority of IDUs shared their needles. HCV genotypes 1 and 2 are the most prevalent in Morocco while genotypes 3, 4 and 5 are less common. Sequence analysis of HCV NS5B region of genotype 2 has shown that HCV subtype 2i is predominant. IDUs exhibited genotypes 1a, 3a and 4 predominantly, as compared to the predominant 1b and 2a/2c genotypes found in general population. The IDUs genotypic profile closely matches the one in other European countries which are invariably speculated as the potential source of currently-circulating genotypes in Moroccan IDUs. On the other hand, HBV genotype D is predominant in Morocco, as this is the major HBV genotype in Mediterranean countries. High circulation of precore and basal core promoter mutants is common in chronic hepatitis B infection in Morocco.
- Sessions: Global Epidemiology of HIV/AIDS, STDs, & STIs | Viral Hepatitis | Veterinary Virology | General Virology and Basic Science | Viral Pathogenesis,Virulence & Latency | Bacterial Virology | Viruses and Diseases
Chair
Shubhada Bopegamage
Slovak Medical University, Slovakia
Co-Chair
Abdelouaheb Benani
Pasteur Institute of Morocco, Morocco
Session Introduction
Shubhada Bopegamage
Slovak Medical University, Slovak Republic
Title: In vivo mouse model for coxsackievirus infections: our story
Time : 10:35-10:55
Biography:
Shubhada Bopegamage is a Virologist. Currently, she is the Head of the Enterovirus Laboratory and the National Reference Center for Identification of Enteroviruses at the Medical Faculty of the Slovak Medical University in Bratislava, Slovakia. Her work is focused on the pathogenesis and diagnosis of Enteroviruses. She received her BSc Microbiology degree from Pune, India and MSc Microbiology degree from Mumbai, India. She got her PhD in Biological Sciences from the Academy of Medical Sciences, Moscow, Russia. She is known in the Enterovirus research, since 2005, for her work on the in vivo experimental coxsackievirus oral infection of mice, and experimental coxsackievirus infection during gravidity. She is involved in research and teaching and has guided several MSc and PhD students. She has Coordinated and has lead several national and international projects as a Principal or Co-investigator
Abstract:
Statement of the Problem: The introduction of cell culture techniques has simplified the work of Virologists. Scientific history shows us, however, that we cannot study a disease completely without using animal models. Though, humans are the natural hosts for coxsackieviruses B (CVB), mice have been shown to be susceptible. In mice, these viruses induce diseases like those which they cause in humans. The experimental animal models allow investigation at a pathophysiological level where the biological picture and functioning is depicted. Though the closest experimental models for the Enteroviruses and human diseases are maybe chimpanzees, the murine models (mice), are one of the best models for studying coxsackieviruses.
Methodology & Theoretical Orientation: We studied genetically different mice strains. Permission for the animal work was obtained from the Ethics Committee of the Slovak Health University and the State Veterinary and Food Control Authority of the Slovak Republic. Mice were infected with different CVB strains and control mice with PBS by oral gavage. Organs were collected from infected mice/groups and control (mock infected) mice/groups at different days post infection. Organs were fixed for histology and immunohistochemistry in formalin and snap frozen and stored at -80ºC for detection of the viral RNA by polymerase chain reaction (PCR).
Findings & Conclusions: Our work showed differences between the oral route and intraperitoneal routes of infection. The differences were mainly in the histopathology of the pancreas, they also depended on the genetic background of the mice. We also showed pancreatitis in challenged pups of mice orally infected dams during gravidity at different times. Our work characterizes and shows the application of our model of the natural ‘oral’ route of infection to understand the pathogenesis of CV infections.