This study aims to conduct pilot surveillance for Nipah virus (NiV), Japanese encephalitis virus (JEV), and Crimean-Congo hemorrhagic fever virus (CCHFV) in East Malaysia, Mexico, Mongolia, and Northern Vietnam. NiV is a highly lethal paramyxovirus that is transmitted from bats and pigs, causing severe encephalitis with a 58% fatality rate. JEV is a flavivirus transmitted by mosquitoes that causes encephalitis in rural and peri-urban Asia, with a 30% fatality rate for symptomatic cases. CCHFV is a bunyavirus transmitted by ticks or infected blood, causing severe hemorrhagic fever with 40% fatality in hospitalized cases. This study will partner with the National Bio and Agro-Defense Facility (NBAF) in attempting to identify and characterize viruses or serological evidence of such viral infections in wildlife and livestock in the four countries.  identify and characterize these viruses in Data from this study may inform future public health interventions and targeted surveillance efforts. 

This study evaluates the novel RABV-H5cow vaccine, a rabies virus (RABV)-based inactivated vaccine designed to protect dairy cattle against highly pathogenic avian influenza (HPAI) H5N1. Previously tested in 12 beef cattle, this study expands the evaluation to 100 dairy cows, divided into six treatment groups with different vaccine combinations. Blood, nasal, and milk samples will be collected over 12 months for the immune and molecular analysis. The immunogenicity will be assessed to measure the antibody response against H5 hemagglutinin and RABV. This immune response will be compared against two USDA-approved adjuvants: MONTANIDETM ISA 201 VG and MONTANIDETM ISA 207 VG. The general health, milk production, and potential vaccine residue in milk will be monitored to ensure safety. This study represents a novel approach to controlling zoonotic influenza using a rabies virus-based vaccine, offering a promising solution for livestock and public health. The vaccine is anticipated to provide dual protection against H5N1 and rabies, benefiting cattle, other ungulates, and even domestic animals. This vaccine has the potential to illicit long-term immunity, similar to the rabies vaccine. The findings from this study may guide the future development of a universal livestock vaccine, while also informing human vaccine applications. 

Often serious viral illnesses are identified via carefully designed molecular laboratory tests.  However, sometimes these carefully designed tests for known pathogens will miss newly identified or rare pathogens.  Through this research project we are working closely with international collaborators and the US Department of Agriculture in seeking to identify such new or rare viruses. Specifically, this study seeks to identify Nipah viruses, Japanese encephalitis viruses, Crimean-Congo Hemorrhagic Fever viruses and other related viruses that might be causing hospitalizations in Malaysia, Mexico, Mongolia, and Vietnam. As we identify such viruses, we will characterize them with a goal of better understanding their epidemiology and developing countermeasures to reduce their future morbidity and mortality.

PI - Gregory Gray

Through the proposed work we seek to better understand the epidemiology of the new canine-like coronavirus, CCoV-HuPn-2018, we recently discovered in a child with pneumonia in Sarawak, Malaysia.  Our interdisciplinary and international team will achieve this by developing molecular and serologic tools to detect the CCoV-HuPn-2018 or CCoV-HuPn-2018 infection. Next, we will use these tools and a cross-sectional study of 200 pneumonia patients and 50 controls to determine how common the virus is among pneumonia patients and the general population in in Sarawak, Malaysia. Finally, we will examine study questionnaire data from study participants to see if we can identify risk factors for CCoV-HuPn-2018 infection.

PI - Gregory Gray

In early 2017, a 5th wave of H7N9 avian influenza (AI) virus infections in man demonstrated the increasing spread of H7N9 virus among poultry in China. In 1997, highly pathogenic H5N1 AI virus outbreaks were reported in Hong Kong and soon became widely spread among poultry in China. It eventually crossed the border into Vietnam and became enzootic among Vietnam’s aquatic birds and poultry. There is growing concern that H7N9 will likewise become enzootic among poultry in Vietnam and cause frequent severe infections in man. Surveillance for AI viruses in Vietnam is very limited.

During the first three weeks of October 2017, Duke, Duke-NUS, and NIVR partnered in conducting a pilot study of aerosol and bird tracheal swab sampling in Hanoi’s largest live bird market (Havi). Ninety percent of 30 four-hour aerosol samples were positive for influenza A by qRT-PCR and these results correlated highly with pooled oral swab cultures from nearby live birds which were 47% qRT-PCR positive. Egg cultures of aerosol and pooled oral swab cultures yielded 26 live viruses, five of which were H5.  In this pilot, there was as yet no evidence of H7 viruses in this market.  Our NIVR collaborators believe this bird market to be at high risk of incursions with H7 viruses from China.

In this proposal, we will conduct a 12-month pilot study of active surveillance for influenza A viruses at Hanoi’s largest bird market and in six live bird markets sites near Vietnam’s northern border with China. Our teams will travel to the live bird market in Hanoi weekly and to six live bird markets in three remote provinces in Vietnam, conducting 10 market visits per month. During these market visits, we will employ novel bioaerosol AI surveillance techniques (less invasive, larger sampling area, more sustainable), along with bird and cage swabbing, as well as collect nasal washes and qualitative survey-based interviews from a convenience sample of poultry workers. Molecular and culture work will be performed at NIVR in Hanoi with close collaboration (periodic visits) from Duke and Duke-NUS scientists. We estimate a 12-month total of 360 bioaerosol samples, 600 bird swab samples, 1200 environmental cage swab samples, and 600 poultry worker nasal wash samples with questionnaires. The samples will be studied with molecular assays, egg or virus culture, and when appropriate by sequencing studies in our Duke One Health laboratory network to determine influenza A type.

PI - Gene Olinger

The Crimean-Congo Hemorrhagic Fever virus (CCHF) is the most widely spread tick-borne pathogen, with cases occurring in Africa, the Middle-East, Eastern Europe as well as Asia. Livestock are amplifying species in CCHF circulation in endemic areas. As a result, individuals caring for or butchering livestock are at higher risk of disease acquisition. Despite CCHF substantial case fatality and broad geographical distribution, there are no specific treatments. Furthermore, the only specific vaccine, an inactivated one, is only approved in Bulgaria due to the associated safety concerns.

Using funds from two Canadian agencies, namely the Department of Foreign Affairs and the International Development Research Center, we are tackling both CCHF surveillance and transmission. First, using a One-Health approach, we are training scientists in West Africa (Mauritania, Ivory Coast) in the detection and sequencing of CCHF isolates from human, livestock and tick samples. Second, we are testing the ability of two DNA vaccines to prevent CCHF infection in livestock. One vaccine encodes for CCHF glycoproteins, while the second vaccine aims at preventing ticks infestation by encoding a gut antigen from ticks.

By building diagnostic capacity for humans, animals and tick vectors, this project will provide a better understanding of CCHF circulation and provide novel CCHF isolate sequences to improve future diagnostic assays and vaccines. Our livestock immunization trial will also provide an affordable means of reducing CCHF transmission to at-risk populations in West Africa.

PI - Dr. Gayani Tillekeratne, Division of Infectious Diseases and the Duke Global Health Institute, Duke University
Dr. Gregory Gray, Division of Infectious Diseases, University of Texas Medical Branch
Dr. Benjamin Anderson, Duke Kunshan University, Kunshan, China

During the last 25 years the world has experienced the emergence and spread of a number of novel respiratory viruses including Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) in 2003 and 2004, and now SARS-CoV-19. Similarly, in 2009, a triple reassortment H1N1 swine-like influenza virus emerged in Mexico, and quickly spread throughout the world. While these novel emergent respiratory viruses are from different families, they have one thing in common, they are thought to have originated in animal and crossed over to infect man. While a variety of risk factors associated with emerging respiratory viruses have been study, the risk factors for specific viruses are complex; however, they tend to be found in densely populated geographic areas of high emerging infectious disease risk and are also often correlated with live animal markets and industrialized farming.

The primary aim of the study is to identify and characterize viruses and determine risk factors associated with pneumonia among patients admitted to participating hospitals in Malaysia, Sri Lanka, Kenya, China, and Vietnam. Teams at collaborating institutions will enroll and collect nasopharyngeal (NP) swab and conduct qualitative surveys to determine risk factors associated with pneumonia. NP swab samples will be studied with molecular assays, virus culture, and when appropriate by sequencing, to characterize viruses. We will also study the laboratory results with their linked questionnaire data with a goal of identifying potential risk factors for pneumonia hospitalization.

PI - Gene Olinger

In resource limited settings, outbreak responders including healthcare providers are particularly vulnerable to infections during infectious disease outbreaks. The high level of casualties amongst healthcare workers ignited debates about whether as caregivers they have a duty to attend victims of lethal epidemics, or if they have a right to stay away from work during such emergencies.

The goal of this project aims at providing guidelines to healthcare workers during infectious disease outbreaks in order to help them deal with potential ethical dilemmas. These guidelines will be designed so that as many patients as possible are given the care they require, whilst securing the safety of healthcare workers and safeguarding the functionality of the healthcare system beyond the contagion.

This work is performed in collaborations with healthcare workers from the Democratic Republic of the Congo which have experienced Crimean-Congo hemorrhagic fever and/or Ebola virus outbreaks. Specifically, we aim to conduct oral interviews with local healthcare providers to fill up a questionnaire that will be used to better understand the perspectives, opportunities, and constraints that they experienced. Their answers will be invaluable to draft recommendations regarding policies to address the ethical dilemmas they may face while minimizing casualties in outbreak responders.

PI - Dr. Gene Olinger

To this day, various communities in Central and West Africa remain isolated from larger urban centers or major trading routes. As such, cases of emerging pathogens in these populations would probably not be reported to local health authorities and thereby increase the risk of pathogen amplification, resulting in larger outbreaks and/or public health emergencies of international concern. Recent outbreaks have also made it clear that inadequate consideration of social, cultural, political, and religious factors in humanitarian responses has fatal consequences regarding the effectiveness and acceptability of response activities.

Working with partners in the Democratic Republic of the Congo, we aim to identify locally-driven solutions to effective surveillance and early detection of pathogens in a range of isolated communities. This approach would be complemented with the identification and establishment of trustful communication channels between select communities and the local health authorities and/or other responders that could be activated during a disease outbreak. This would be accomplished using a wide array of methods such as community-based research groups, interviews with key stakeholders, as well as participant observations.

Overall, this approach would create multi-level partnerships between remote communities and local and external health authorities, allowing surveillance plans to be supported and acted upon.

PI - Dr. Gene Olinger

Each year, the World Health Organization (WHO) produces a list of priority pathogens, for which scientific resources should be invested towards in the upcoming year. These pathogens usually exhibit a higher outbreak potential and do not have readily available medical countermeasures. In Central and West Africa, more often than not, only limited or outdated data is available on the prevalence of these pathogens.

We are therefore working with local institutions in Africa to provide them with training and diagnostic capacities. Depending on the country, the targeted priority pathogens include Crimean-Congo hemorrhagic fever (CCHF) virus, Ebola virus, Marburg virus, Lassa virus, Middle East respiratory syndrome coronavirus (MERS-CoV), severe acute respiratory syndrome (SARS) virus, Nipah virus and henipaviral diseases, Rift Valley Fever (RVF) virus and Zika virus. Monkeypox and Leptospirosis, for which the WHO also noted that a more comprehensive surveillance program is needed, will also be included when possible.

In addition to training local individuals, the above research-based training program will provide information on the prevalence of various infectious threats from the WHO list of priority pathogens in various locations in Central and West Africa.

PI - Dr. Toh Teck Hock, Pediatrician and Head of Clinical Research Centre, Sibu Hospital, Sibu, Sarawak, Malaysia

Co-PI - Dr. Gregory Gray, Division of Infectious Diseases, University of Texas Medical Branch

Southeast Asia has been identified as a hotspot for emerging infectious diseases including arboviruses such as Japanese encephalitis, dengue, chikungunya, and Zika virus. In Malaysia, dengue is commonly reported, affecting roughly 50-100 million people with a combined death toll estimated to be 24,000 per year globally. While the capacity for arbovirus diagnostics is good in some parts of the country, the equatorial state of Sarawak, Malaysia, located on the island of Borneo, only has the capacity for rapid combo tests (RCT) which are exclusively used in dengue detection and the ability to surveil for other arboviruses are not routinely available.

The T-COR 8 Real-time PCR Thermocycler (Tetracore Industries, Rockville, MD, USA) increases the span of surveillance possible for clinicians in Sarawak due to the breadth of its diagnostic capability, as well as its portability. Therefore, the overall goals of this study are to 1) evaluate the clinical value of the T-COR 8 multiplexing platform for detecting dengue, chikungunya and Zika viruses; and 2) determine the prevalence of dengue, chikungunya and Zika virus infections among patients with dengue-like infection symptoms in medical facilities in Sarawak. Teams at two collaborating institutions, Sibu and Kapit medical facilities, in Sarawak, Malaysia will enroll and collect serum, gingival crevicular fluid, and urine samples and conduct qualitative surveys to determine risk factors associated with arboviruses. An aliquot of serum, gingival crevicular fluid, and urine samples will be run on the T-COR 8 at Sibu and Kapit facilities. A second paired aliquot of sera will be studied with US CDC gold standard molecular assays at Sibu to determine the sensitivity and specificity of the Tetracore results. A third paired aliquot of sera will be shipped to Tetracore Inc. for antibody assays. UTMB staff will provide guidance on laboratory assays and will provide statistical support to determine risk factors associated with arbovirus infection.

PI - Miguel M. Cabada, MD, MSc, University of Texas Medical Branch, Director of UPCH-UTMB Research Center in Cusco, Peru

Dr. Cabada is the Principal Investigator for a recently awarded Tropical Medicine Research Centers (TMRC) grant from NIAID titled: One-Health Approach to Study Human Fasciola hepatica Transmission and Inform Strategic Control. The TMRC are very competitive cooperative agreement grants available every 5 years to advance NIAID’s global research effort by targeting research endeavors in low and middle-income countries and create and sustain in-country research capacity. Fasciola is a liver fluke with a complex lifecycle that is influenced by human behavior and environmental conditions and involves snails, livestock, and humans. The fluke disproportionately affects children in endemic countries causing anemia, malnutrition, and liver disease. A one health approach modeling all these variables will be able to identify limiting points in transmission that can be intervened on to control the infection. Dr. Cabada will be working in Peru with investigators from UTMB, Oregon Health and Science University, Universidad Peruana Cayetano Heredia, and Universidad Nacional de Cajamarca disentangling the factors that determine the transmission of Fasciola to humans and creating new diagnostic and surveillance tools to control the infection.

Image 1 - Above: Environmental conditions that allow the maintenance of Fasciola cycle in Cusco

Image 2 - Collection of snails in the field to evaluate infection rates

Image 3 - Snail colonies and metacercariae that emerged from infected snails.

Summary

Mongolia has some of the highest ambient particulate matter (PM) concentrations in the world and frequently suffer epidemics of viral respiratory disease. There is some evidence to suggest that PM exposure may weaken the immune defense against respiratory viral infection thereby enhancing incidence and severity of infection but this has not been well studied. In collaboration with the Mongolian National University of Medical Sciences (MNUMS) and Duke University, we will test the hypothesis that PM exposure weakens the immune defense against respiratory viral infection through a main study and two pilot studies. Specifically we first aim to characterize the relationship between PM exposure and respiratory viral infection through a time-series analysis of weekly influenza-like illness rates in association with PM levels in two major Mongolian cities, Ulaanbaatar (UB) and Darkhan City (DC).

Secondly, we aim to examine immunological and viral responses to personal PM exposure by following 100 nonsmoking adults in both UB and DC every 6 months for three years and collect questionnaire data and biological specimens for lung function, inflammatory responses, and blood pressure. Air samplers will also be employed to measure both PM exposure and to detect airborne viruses.

The third aim of this project is to define the direct effects of PM exposure on epithelial integrity and immune signaling and the subsequent impact on anti-viral responses. This will be done through cell culture studies directly linked to aim 2 subjects (as cell donors).

Lastly, in the two pilot studies, we will explore the effects of PM and heavy metals in PM on pulmonary tuberculosis and bone mineral loss, respectively, in family members of the subjects enrolled for Aim 2. UTMB staff will provide guidance, training, and oversight for the measurement of viruses in bioaerosol samples collected in houses for Aim 2. UTMB Positive bioaerosol samples may also be transferred to UTMB for further characterization studies by UTMB staff.

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PI - Gregory C. Gray, MD, MPH, University of Texas Medical Branch

In this 5-year, One Health-oriented, prospective research project we will study 16 US or Mexican livestock farms (pigs, cattle, or poultry) for SARS-CoV-2 and other circulating coronaviruses using our pan-species coronavirus assay. Questionnaire data (farm and livestock workers) and samples will be collected every four months for one year from the farm environment, the farm livestock, and the livestock workers. In between the four farm visits, we will use postage-paid sample kits to collect and ship nasal/oral swabs from livestock or livestock workers with signs of respiratory illness to UTMB. In year 3 of the study, we will field-test on farms a pan-coronavirus assay developed by GeneCapture, Inc. We will intensively study novel coronaviruses with culture, full genome assembly and reverse genetics. Overall, this study aims to develop effective and cost-efficient methods for the rapid detection of SARS-CoV-2 and other coronaviruses on livestock farms. Summary surveillance data and novel pre-pandemic coronavirus detections will be shared with the National Animal Health Laboratory Network for further assessment. We expect to demonstrate that this rather low cost, One Health surveillance approach to farm biosecurity will serve as a model for pre-pandemic pathogen detection and have potential for widespread adoption. Additionally, in support of this work, we will offer practical virology training to up to 40 minority undergraduate students from Galveston College over the five years of research.

PI - Gregory C. Gray, MD, MPH, University of Texas Medical Branch

Since the start of the 20th century, humanity has experienced five waves of influenza pandemics. The large diversity of influenza viruses in animal reservoirs portends a constant risk of the next inevitable pandemic. Current influenza vaccines are designed to protect against seasonal influenza but leave individuals largely susceptible against heterosubtypic influenza viruses. The development of a new vaccine which can greatly expand the breadth of immunity to future pandemic strains would have the potential to save many lives and decrease global morbidity by a significant amount.

The purpose of this project is to perform a Phase II clinical trial to evaluate the efficacy and safety of the BPL-1357 vaccine in the setting of a healthy volunteer influenza human challenge study. The study will be sponsored and guided by NIAID but performed by UTMB’s research team. The primary hypothesis is that IM (intramuscular) and IN (intranasal) BPL-1357 will be safe and offer protection against mild-moderate influenza disease (MMID) caused by H1N1 influenza challenge compared to placebo.

The primary objectives of this study are to measure the efficacy of BPL-1357, given IM or IN, in preventing MMID compared to placebo; to assess the safety of BPL-1357, given IM or IN in 2 doses 28 days apart, followed by viral challenge with H1N1. The challenge will be with a human 2015 seasonal-like H1N1 strain against which participants are likely to have partial immunity. The study population includes healthy male and female volunteers, ³ 18 to £ 55 years of age. The 120 participants are divided into three study arms (placebo, intranasal vaccine, and intramuscular vaccine).

This [KB1] is a randomized, double-blinded, single-center, phase 2 clinical trial of BPL-1357 administered intranasal or intramuscular in 2 doses 28 days apart compared to a single dose of placebo given intramuscular, followed by viral challenge with H1N1. Participants will be randomized to one of three groups for treatment assignment. This study has two phases: a vaccination phase (Phase A) and a challenge phase (Phase B). On enrollment participants will be randomized 1:1:1 to three groups.

PI - Gregory C. Gray, MD, MPH, University of Texas Medical Branch

The UTMB One Health Laboratory will partner with the life science tools company InDevR Inc. (Boulder, CO) in developing and testing a unique serologic assay to detect previous infections from three different influenza viruses: Influenza A (H5N1), Influenza C, and Influenza D. The new diagnostic will be based upon InDevR Inc's successful microarray platform (VaxArray) and available for use during the current H5N1 national epidemic. The species-agnostic microarray will be able to rapidly screen relatively large numbers of specimens at relatively low cost to help farmers, zoo managers, and public health officials determine which animals and animal workers need to receive periodic vaccines against highly pathogenic H5N1 viruses.

PI - Gregory C. Gray, MD, MPH, University of Texas Medical Branch

In partnership with the USDA-ARS, we will leverage our extensive network of national and international research partnerships in the United States and in a number of geographically diverse countries including but not limited to: Vietnam, Pakistan, North Africa, Mexico, Mongolia and elsewhere in performing One Health-related research for potential spillover events. Samples for humans, animals, and the environment in will be interrogated for evidence of spillover pathogens, especially novel respiratory viruses, Japanese encephalitis virus, Nipah and related viruses, Crimean Congo hemorrhagic fever virus and novel tick-borne disease pathogens. Where feasible isolates will be recovered. This effort has the potential to provide critical data on the frequency of spillovers and the collection of samples that can be utilized for the testing of diagnostics and potentially the isolation of agents of interest.