Eastern PA Branch-ASM, 41st Annual Symposium, November 17, 2011

1. The Galveston
National
Laboratory, a The Galveston National Laboratory
national resource
for emerging
diseases and
bioterrorism
threats
Dr. Jim LeDuc
Director
Presented at the 41st Annual Symposium
―Global Movement of Infectious Pathogens and Improved Laboratory
Detection‖
Eastern PA Branch-American Society for Microbiology
November 17, 2011
Thomas Jefferson University, Philadelphia

2. What are the next emerging virus
diseases to threaten the USA?
• Objectives for today:
– Review some of virus diseases of concern as
potentially ―emerging‖
– Introduce the Galveston National Laboratory
as a new national resource to address
emerging infectious diseases
– Provide examples of what’s already underway
at the GNL to address EIDs

3. GNL Mission
• The mission of the GNL is to assist
the NIAID and the nation by
conducting basic and applied
research designed to improve the
prevention, diagnosis and treatment
of naturally emerging and
purposefully disseminated infectious
diseases.

4. Building Section
Galveston National
Laboratory
BSL3 BSL2
BSL2 BSL2
ABSL3
Ike
Surge BSL4

5. © Hedrich Blessing Photography 2009

6. Services Divisions
Services available to researchers in biodefense
and emerging infectious diseases:
 Aerobiology
 Assay Development
 Experimental Pathology
 Imaging
 Immunology (currently inactive)
 Insectaries Services
 Preclinical Studies

7. Basic
Product Development
Research
NIH R01 and
other Funded Proof of Concept
Research Concept Validation
Pre-clinical/
GLP Studies
Human
Animal Rule
Clinical Trials
Phase I: FDA
Safety Restricted Use
Phase II: Approval
GNL Expanded Safety
Phase III:
UTMB Efficacy
FDA
Licensure

8. West Nile Virus in the United States 1999-2008

9. Influenza A Viruses
• Human, Avian, Equine, & Swine Influenza viruses
• Influenza A, B and C
– B & C viruses infect humans only
• A viruses – ordinary flu & all pandemics
– Subtyped by surface glycoproteins; change =
Antigenic Shift
– 16 hemagglutinins (HA) and 9 neuraminidases (NA)
– Current human subtypes: H1N1, H3N2 NA
HA
www.pandemicflu.gov www.cdc.gov/flu

10. Emergence of Influenza A Viruses in Humans
1998 1999
*Avian H9* 2003
Influenza
Viruses
H5* 1997 2003-
2011
H7* 1980 1996 2002
2003
2004
H1
H3
H2
H1 1977
1915 1925 1935 1945 1955 1965 1975 1985 1995 200
1918 1957 1968
Spanish Asian Hong
Flu H1N1 Flu H2N2 Kong
Flu H3N2
www.pandemicflu.gov www.cdc.gov/flu

11. Cases of Avian influenza in People
Year Strain Country No. No.
confirmed confirmed
human cases human
deaths
1997 A/H5N1 Hong Kong 18 6
1999 A/H9N2* Hong Kong 2 0
2003 A/H5N1 Hong Kong 2 1
2003 A/H9N2* Hong Kong 1 0
2003 A/H7N3 Canada 2 0
2003 A/H7N7 Netherlands 84 1
2003- A/H5N1* Viet Nam, 569 334 (59%)
2011 Cambodia*, China,
(Nov11) *WHO Bangladesh*, Thailand
vaccine in Azerbaijan, Djibouti,
Egypt**, Indonesia**,
prep.
Iraq, Myanmar,
Thailand, Turkey,
www.pandemicflu.gov www.cdc.gov/flu
Nigeria, Pakistan, CDC, WHO,

12. Host and lineage origins for the gene segments of the 2009 A(H1N1) virus:
PB2, polymerase basic 2; PB1, polymerase basic 1; PA, polymerase acidic; HA,
hemagglutinin; NP, nucleoprotein; NA, neuraminidase; M, matrix gene; NS,
nonstructural gene
R. J. Garten et al., Science 325, 197 -201 (2009)
Published by AAAS
www.pandemicflu.gov www.cdc.gov/flu

13. Influenza activities at the GNL
• Evaluation of candidate broadly cross
protective vaccines
• Testing of antiviral compounds for
treatment of avian influenza
• High through-put screening of clinical
samples during H1N1 outbreak
• Reagent production for WHO

14. Nipah virus disease
• First recognized in 1998 during outbreak in
pigs in Malaysia
• Annual outbreaks in humans in
Bangladesh
• High mortality rate; serious sequelae
• Potential for person-to-person
transmission (so far not efficient)
• Animal reservoir host (fruit bats)

15. Breaking the Chain in Bangladesh
Science 4 March 2011:
vol. 331 no. 6021 1128-1131

16. Pteropus Bats, suspect reservoir of
Nipah virus

17. Nipah site in Bangladesh

18. Nipah investigations at GNL
• Development and validation of NHP model
for Nipah infection
• Evaluation of monoclonal antibodies for
treatment of Nipah infection
• Basic replication studies
• Vaccine efficacy studies in NHP

19. Rift Valley Fever
History
Kenya, Lake Naivasha 1910 Montgomery
Kenya, Lake Naivasha 1930, 33 Daubney et al
Uganda, Semliki Forest 1948 Smithburn et al
South Africa 1950-1 Alexander & Dickson
1953
1955-56
South Africa 1975 Van Velden et al
Egypt 1977 Meegan et al
Mauritania 1987 Digoutte et al
East Africa 1997-98 MMWR
Arabian Peninsula 2000 MMWR; Madani et al
East Africa 2006-07 MMWR, Bird et al, 2008
Madagascar 2008- Andriamandimby et al,
South Africa 2008-
2009 NIV Bulletin, 2010
2010
2009

20. RVF
Epizootiology/Epidemiology
• Primary vector maintenance
– Pool breeding Aedes spp. maintain via transovarial
transmission (putative)
– Can maintain in a silent manner during extended dry
periods
• Secondary vector species
– Large populations during periodic/cyclic rainy seasons
– Initial amplification in vertebrate hosts
– High viremia levels in many species of vertebrate
hosts overcome poor vector potential of secondary
vectors

21. RVF
Egypt 1977-1978
Governorate Sheep Cattle Humans
Sharqiya 52 36 15
Asyut 40 27 19
Qena 38 21 5
Qalubiya 37 15 6
Aswan 33 57 6
Giza 24 19 3
Fayum 13 0 3
Minya 9 5 4
Daqahliya 3 0.4 1
Baheira 2 1 1
Kafr el Sheikh 0 0 3
Minufyia 0 0.8 2
Beni Suef 0 0 1

22. Summary of data implicating various animals and
humans as amplifying hosts or reservoirs of RVF virus
(Meegan, 1981)
Host Viremia Viremia
(peak (length in
titer/ml) days)
Sheep 1010 2-5
Cattle 109 2-5
Human 108 3-10
Goat 108 2-5
Rattus sp. < 103 <2
Arvicanthus sp. < 103 <2
Acomys sp. < 103 <2
Mus sp. < 103 <2
D EC
2010

24. Effect of Rainfall on RVF
Outbreaks

25. Saudi Arabia
Index Case
Week 1
Week 2
Week 3
Week 4
Week 5
Jizan
0 25 50 Yemen
kilometers

26. Hospitalized cases of Rift Valley
Fever by week, Saudi Arabia,
August 27, 2000 – November 18,
2000
100
N=678
80
60
Number of
RVF Cases
40
20
0
0 1 2 3 4 5 6 7 8 9 10 11 12
Week of Onset

27. Entomological Investigations
• Two major species identified
–Aedes vexans arabiensis
–Culex tritaeniorrhyncus
• Virus isolates from both species
• Transmission from both species
• Some biting preference data
–Human biting preference relatively high

28. Epidemic Curve (MMWR)

29. Adjoining
Countries
(WHO)

31. Vector Competence
North American mosquitoes
following Oral infection (Gargan et al, 1988;Turell et al, 1988)
Species Infection (%) Transmission (%)
Cu. tarsalis 88 36
Ae. canadensis 96 54
Ae. taeniorrhyncus 85 42
Ae. triseriatus 83 36
Ae. albopictus 89 12

32. Vaccines
• Veterinary Vaccines
– Smithburn neurotropic strain (SNS)
– Killed products using formalin inactivated cell culture
antigens
– New generation:
• MP12
• Strain 13
• Reverse genetics: NSS and NSM deletions
• Human Vaccines (IND only)
– Killed cell culture product (formalin)—USAMRIID
–MP12—attenuated (mutagen passage) live
product--USAMRIID

33. RVF studies at GNL
• Advanced development of MP12 live,
attenuated vaccine for RVF virus
– Pilot lot production with commercial partner
– Animal model development and
standardization
– Vaccine efficacy testing in animal models
• Basic studies on viral replication and host
cell targeting

34. Chikungunya Virus

35. 2007
2005
2008
before
1950`s 2004 2007
2007 2008
2005 Countries endemic for ECSA genotype
Countries endemic for Asian genotype
Countries endemic for WA genotype
2005 Regions invaded by CHIKV strains of IOL
during 2004-2008
2006 Reconstructed movement of CHIKV during
2004-2008 outbreaks
Introduction of progenitor of Asian Genotype
before 1950`s

36. E1-226A
E1-226V
IOL
E1-98A
E1-98T
ECSA
Asian
WA

37. Global destitution of Ae. albopictus
Map showing the native habitat (blue) and recent spread (green) of the Aedes
albopictus mosquito.
Downloaded from http://afludiary.blogspot.com/2010/06/mmwr-travel-associated-
dengue.html

38. Distribution of Ae. albopictus in the US.
Ae. albopictus in USA as of 2001. Adapted form (Benedict et al., 2007)

39. Distribution of Ae. albopictus in Europe.
Ae. albopictus in Europe per province, as of January 2007. Adapted form
(Scholte and Schaffner, 2007)

40. CHIK in Yemen?
Health Ministry warns of fatal illness spreading from Yemen
Sunday, 20 March 2011 14:52 JEDDAH/SANAA:
Many people who live on the Saudi-Yemen border fear that a fatal disease
that has been responsible for dozens of deaths in Yemen’s western
coastal area could cross into the Kingdom.
At least 65 deaths were reported in Yemen’s western coastal province of
Hodeidah.
The disease is thought to be Chikungunya, though some medical officers
dispute it. Symptoms of chikungunya include kidney failure, high
temperature, diarrhea and vomiting.
Dr. Abdul Hakim Al-Khohlaini, manager of the epidemiological
surveillance department at the Yemeni Ministry of Health, said that
samples sent to Cairo for tests showed that the disease is
chikungunya. He added that local doctors have also found other
diseases in the area.
“Forty-five percent of the cases are Chikungunya, 10 percent malaria
and the rest dengue fever,” he said.

41. •Tick-borne zoonosis; discovered 1944 in Crimean peninsula,
isolated from over 30 countries
•Causative agent Crimean-Congo hemorrhagic fever virus (CCHFV)
•Hard ticks of the genus Hyalomma are the main vector
•Severe hemorrhagic syndrome in humans; up to 70% mortality
•ssRNA- , Bunyaviridae, Nairovirus (7 species, 34 strains)
•Genome analysis indicated considerable evolution and
high diversity of CCHFV strains (array of genotypes)
•Different genotypes might cause different severities of
disease
•Experimental work with CCHFV requires a BSL-4 laboratory.
•No vaccine, Ribavirin treatment uncertain efficiency
•Outbreaks are usually sporadic with only a few cases but
recent bigger outbreaks in Balkan and Southwest Asia
demonstrated imminent public health threat

44. • Outbreaks are usually sporadic with only a
few cases but recent bigger outbreaks in
Balkan and Southwest Asia demonstrated
imminent public health threat.
• CCHFV first detected in Turkey in 2002;
since then >4000 cases.
• More here to come!

45. 1. To study the virulence of different CCHFV strains
currently circulating in the Balkan, Caucasus and
Turkey in in vitro an in vivo models.
2. To establish a tick in vivo feeding assay in which
the host-tick-interface can be studied.
3. Use the in vitro feeding assay to study
 the vector capacity of and North American tick species
to CCHFV transmission
 If viral attenuation is correlated with tick vector switch
(Hyalomma spp. vs Rhipicephalus bursa)

46. Novel Bunyavirus in China
 Designated SFTS virus (Severe Fever with
Thrombocytopenia Syndrome)
 First isolated from acutely ill patient
 Found in rural areas of Hubei and Henan provinces; 96%
of cases between May-July
 Ages 39-83 y.o., 56% female, 97% working farmers
 30% case fatality rate initially; 12% (21/171 confirmed
cases) on further investigation
 Genus Phlebovirus, perhaps new subgroup
 35 RT-PCR confirmed cases all seroconverted
 Suspect Tick transmission

47. Galveston National Laboratory

48. The disease is a severe hemorrhagic fever with
very high fatality
Clinical signs and symptoms
• Fever
• Leukopenia (low white blood
cells)
• Thrombocytopenia (low platelet
count)
• Gastrointestinal disorders
(vomiting and diarrhea)
• Hemorrhaging
• Multiple organ failure
• Death (on average 12% fatality
rate and as high 30% in some
areas)
Galveston National Laboratory

49. Clinical Symptoms of Hospitalized Patients with Laboratory-Confirmed SFTS.
Yu X et al. N Engl J Med 2011. DOI:
10.1056/NEJMoa1010095

50. The environment of a recovered patient is infested with ticks
A farmer and his house The famer cultivated mushrooms Auricularia
(Auricularia auricula) outside his house auricula
The yard of the farmer Collecting ticks from a dog by the Collecting ticks from a dog by the
house owner owner
Galveston National Laboratory

51. Geographic Distribution of SFTS in China.
Yu X et al. N Engl J Med 2011. DOI: 10.1056/NEJMoa1010095

52. Phylogenetic Analysis of SFTS Bunyavirus and Other Phleboviruses.
Yu X et al. N Engl J Med 2011. DOI: 10.1056/NEJMoa1010095

53. The discovery of the severe fever with thrombocytopenia
syndrome virus
Xue-jie Yu, M.D., Ph.D. Patricia V. Aguilar, Ph.D.
Professor Assistant Professor
Department of Pathology Department of Pathology
Center for Biodefence and Emerging Infectious Diseases Center for Biodefense and Emerging Infectious Diseases
Sealy Center for Vaccine Development Institute for Human Infectious and Immunity
Galveston National Laboratory

54. And Finally….A new Flavivirus
from China
―Duck Egg-Drop Syndrome Caused by BYD
virus, a New Tembusu-related Flavivirus‖
by J. Su et al. PLoS One, March 2011.
• Infection completely eliminated duck reproduction on
some farms; duck mortality 5-15%;
• Baiyangdian (BYD) virus isolated from brain and ovary of
affected ducks
• Shown to be a flavivirus closely related to Tembusu and
Sitiawan & Bagaza viruses, some known to cause
disease in humans and animals.

55. In Conclusion
• There’s no shortage of emerging virus diseases
• Many are zoonoses, often vector-borne
• Expanding sequence databases and improved
techniques allow rapid recognition and
characterization of new viruses
• The GNL is available as a national resource to
assist in basic and translational research on
emerging viruses
• www.UTMB.edu/gnl

56. Key Contacts
Director:
James W. LeDuc, Ph.D.
Scientific Director:
Scott C. Weaver, Ph.D.
Associate Director for Research &
Operations:
Joan Nichols, Ph.D.
Assistant Director:
Andrew G. McNees, Ph.D.
Director of Containment
Operations:
Thomas G. Ksiazek, D.V.M.,
Ph.D.
Director of Strategic Initiatives: Galveston National Lab • Main: 409-266-6500
Alisha Prather www.utmb.edu/gnl
Director of Information Technology:
Bryan Fisher
External Research Coordinator:
Efthalia L. Bataki, Ph.D.