- 1.1 GENERAL INFORMATION CONFERENCE LOCATION Hilo Hawaiian Hotel
71 Banyan Dr Hilo, HI 96720 Telephone: 808-935-9361 USA/Canada Toll
Free - 1 (800) 367-5004 Fax: 1 (808) 969-6472 EMERGENCY The nearest
hospital to the Hilo Hawaiian Hotel is: Hilo Medical Center 1190
Waianuenue Ave Hilo, HI 96720 Telephone: (808) 974-4700 PARKING
Complimentary parking is available for IWOP-11 participants at the
Hilo Hawaiian Hotel REGISTRATION AND MESSAGE BOARD The registration
desk will be located in the Hilo Hawaiian Hotel Foyer, and open
during the following times: Sunday, August 1, 2010; 4:00 6:00pm
Monday, August 2, 2010; 7:30am 4:00pm Tuesday, August 3, 2010; 7:30
10:00am Wednesday, August 4, 2010; 7:30am 4:00pm Thursday, August
5, 2010; 7:30am 3:00pm There will be a board available for posting
messages in the registration area. HELP DESK Members of the Local
Arrangements Committee and the University of Hawaii at Hilo
Conference Center Staff can be contacted at the Registration Desk
in the Hilo Hawaii Hotel, Ground Level Foyer. SPEAKER READY ROOM
The Moku Ola 1 Room will be set up for platform presentations. In
an effort to provide you with the best presentation experience and
ensure that your needs are met, we would like to outline the
presentation review guidelines, laptop specifications, and
audiovisual equipment with you. PRESENTATION REVIEW PROCESS We have
created a review process to eliminate potential incompatibilities
between your personal software versus the presentation software, as
well as give you the opportunity to to rehearse your presentation
with our audio-visual technician, Robin Black, once you arrive
onsite.
2. 2 If you will be using PowerPoint for your presentations,
please have a copy of your file on USB/CD-Rom upon registration in
the Foyer of the ground level of the Hilo Hawaiian Hotel. Once you
have registered, please make an appointment with Robin Kealoha
Black, who will be reviewing all talks to ensure a smooth program.
MAC Users: If you have created your presentation on a MAC, please
visit our website for guidelines in order to have a successful
presentation: http://uhhconferencecenter.com/av-
guidelines-for-mac-users POSTER BOARDS & SESSION INFO Poster
boards are 8' x 4'; each poster presentation should not be larger
than 4' x 4' Poster Session A, #s 1-18; Monday, August 2, 2010;
10:30am 12:00pm Set-up: Sunday, August 1, 2010; 4:00 6:00pm
Take-down: Monday, August 2, 2010; 5:00 5:30pm** Poster Session B,
#s 19 38; Wednesday, August 4, 2010; 10:30 12:00pm Set-up: Monday,
August 2, 2010; 5:30 6:00pm Take-down: Thursday, August 5, 2010;
3:00 4:30pm** **Please Note: Any posters left up after the
designated take-down periods will be discarded. WEATHER The summer
temperatures on the Big Island of Hawaii normally range from 77--80
F (25-- 26.7 C) during the day and from 61--64 F (16.1--17.8C) at
night. Hilo is on the windward side of the island where rainfall
may be 5--10 times higher than on the leeward side. BE PREPARED FOR
ENJOYING IWOP-11 AND OTHER ACTIVITIES IN RAIN. The summer
temperatures on the Big Island of Hawaii normally range from 77--80
F (25-- CURRENCY Foreign currency exchanges: Hilo Hawaiian Hotel
(no currency exchange available; ATM machine in convenience store);
Bank of Hawaii, Kawili Street (Yen, Euro Bank notes, Australian,
Canadian, British, Hong Kong, New Zealand, Singapore); Naniloa
Hotel (Yen exchange only, ATM machine available at various
locations in town. 3. 3 PROGRAM SCHEDULE IN THIS BOOKLET, THE CODE
INDICATED AFTER THE PRESENTATION TIME CORRELATES WITH THE CODE
ASSIGNED TO EACH ABSTRACT. THE ABSTRACTS ARE FOUND AFTER THE
PROGRAM SCHEDULE AND ARE ARRANGED CHRONOLOGICALLY (IN THE ORDER OF
PRESENTATION) OR THE POSTERBOARD ASSIGNMENT. AN INDEX OF AUTHORS IS
FOUND AT THE END OF THE BOOKLET. SUNDAY, AUGUST 1, 2010 PM 4:00
REGISTRATION OPENS FOYER, LOWER LEVEL 4:00 SET-UP OF POSTERS #1-18
LOWER LEVEL, MOKU OLA II 6:00 - 8:00 OPENING RECEPTION, WELCOME
& ANNOUNCEMENTS OPEN TO ALL REGISTRANTS LOWER LEVEL [MOKU OLA
BALLROOM AND GARDENS] MONDAY, AUG 2. 2010 AM 7:30 REGISTRATION
FOYER, LOWER LEVEL PLATFORM SESSION I MOKU OLA I CO-CHAIRS -
CCILE-MARIE ALIOUAT-DENIS, RUSSEL HAYMAN 8:00 PL1 The Biochemistry
and Genetics of Mouse Resistance to Toxoplasma gondii: the Role of
Immunity-Related GTPases (IRG proteins). J. P. HUNN, A. KHAMINETS,
J. LILUE, B. MUELLER, N. PAPIC, N. PAWLOWSKI, T. STEINFELDT, J. C.
HOWARD; Institute for Genetics, University of Cologne, Cologne,
NRW, Germany. 8:20 PL2 Involvement of Integrin Alpha 2 (ITGA2) in
Cryptosporidium parvum Infection. 4. 4 HAILI ZHANG, FENGGUANG GUO,
GUAN ZHU; Department of Veterinary Pathobiology, Texas A&M
University, TX, USA. 8:40 PL3 Lactosylceramide Mediates
Pneumocystis Beta-Glucan Activation of the Human IL-23-IL-17 Axis.
EVA M CARMONA, ANDREW H LIMPER. Thoracic Diseases Research Unit,
Mayo Clinic, Rochester, MN, USA. 9:00 PL4 Pneumocystis carinii
Lanosterol Synthase Requires Saccharomyces cerevisiae 3-
Ketoreductase for Localization to Lipid Particles in Yeast. TIFFANY
M. JOFFRION,1 MELANIE T. CUSHION1,2 . 1 University of Cincinnati
College of Medicine, Cincinnati, OH 45267, USA; 2 Veterans
Administration Medical Center, Cincinnati, OH 45220, USA. 9:20 PL5
Characterization of Lanosterol Synthase (Erg7) from Pneumocystis
jirovecii. THOMAS M. SESTERHENN,1 A. GEORGE SMULIAN,1,2 MELANIE T.
CUSHION1,2 ; 1 University of Cincinnati College of Medicine,
Cincinnati, OH, USA, 2 Cincinnnati Veterans Affairs Medical Center,
Cincinnati, OH, USA. 9:40 PL6 Bone Marrow Failure and Immune
Responses to Pulmonary Pneumocystis (PC) Infection. DAVID TAYLOR,
MICHELLE WILKISON, NICOLE MEISSNER, Department of Veterinary
Molecular Biology, Montana State University, Bozeman, MT 59718, USA
10:00 BREAK 10:30 POSTER SESSION A - NUMBERS 1-18 MOKU OLA II PO1
Cell Wall Assembly Components of Pneumocystis carinii Exhibit
Unique Activity in Response to Alterations in Environmental
Temperature. DEANNE M. HEBRINK1 , THEODORE J. KOTTOM1 , ANDREW H.
LIMPER1 ; 1 Thoracic Diseases Research Unit, Mayo Clinic,
Rochester, MN, USA. PO2 Influence of Climate and Ambient Air
Pollutants on Pneumocystis jirovecii Pneumonia (PcP) Hospital
Admission and on Antibody Responses to Major Surface Glycoprotein
(Msg) in HIV-Infected Patients from San Francisco. K. DJAWE,1 L.
HUANG,2 K.R. DALY,1 , L. LEVIN,1 J. KOCH1 , A. SCHWARTZMAN,2 S.
FONG,2 B. ROTH,2 , A. SUBRAMANIAN,2 K. GRIECO,2 L. JARLSBERG,2 P.D.
WALZER1 . 1 VAMC and U. Cincinnati, Cincinnati, OH, USA, 2 San
Francisco General Hospital (SFGH) and U. California SF, San
Francisco, CA, USA. PO3 Pneumocystis jirovecii Colonization in
Patients Undergoing Treatment with the TNF- Antagonist Agent
Infliximab. RUBN MORILLA1 , ISABEL MARTN- GARRIDO1 , GUSTAVO
WISSMANN2 , VICENTE FRIAZA1 , NIEVES RESPALDIZA1 , RAFAEL TERAN1 ,
MARIA T. MARTINEZ-RISQUEZ1 , 5. 5 ELENA CAMPANO1 , FRANCISCO J.
MEDRANO1 , CARMEN DE LA HORRA1 , JOS M. VARELA1 , JUAN POVEDANO3 ,
ENRIQUE J. CALDERN1 ; 1 Instituto de Biomedicina de Sevilla and
CIBER de Epidemiologia y Salud Pblica. Service of Internal Medicine
Virgen del Rocio University Hospital, Seville, Spain; 2 Grupo de
Estudos em Pneumocystis, Servio de Infectologia, Hospital de
Clnicas de Porto Alegre, Brasil; 3 Service of Rheumatology, Virgen
del Roco University Hospital, Seville, Spain PO4 Dihydropteroate
Synthase Mutations in Pneumocystis jirovecii Isolated from Patients
with Pneumocystis Pneumonia in Brazil. GUSTAVO WISSMANN1 , ROSECLER
B. MENDES1 , VICENTE FRIAZA2 , ANDR L. MLLER1 , RUBEN MORILLA2 ,
CARMEN DE LA HORRA2 , LUCIANO Z. GOLDANI1 , ENRIQUE J. CALDERN2 ; 1
Grupo de Estudos em Pneumocystis, Servio de Infectologia, Hospital
de Clnicas de Porto Alegre, Brasil; 2 Instituto de Biomedicina de
Sevilla and CIBER de Epidemiologia y Salud Pblica. Virgen del Rocio
University Hospital, Seville, Spain. PO5 DHPS Mutations in
Pneumocystis jirovecii Isolated from Patients with PcP in Santiago,
Chile. CAROLINA A. PONCE,1 MAGALI CHABE,2 CLAUDIO GEORGE,1
ALEJANDRA CARDENAS,1 LUISA DURAN;2 JULIA GUERRERO,1 LAURENCE
HUANG,3 ROBERT F. MILLER,4 SERGIO L. VARGAS1 ; 1 University of
Chile School of Medicine, Santiago, Chile, 2 Institute Pasteur du
Lille, France, 3 University of California San Francisco, San
Francisco, CA, USA, 4 University College London, UK. PO6
Characterization of Mouse Alveolar Macrophage Binding to
Pneumocystis murina Cyst and Trophic Forms. A.D. ASHBAUGH, M.J.
LINKE, M.T. CUSHION; Veterans Affairs Medical Center, Cincinnati,
OH, and University of Cincinnati College of Medicine, Cincinnati,
OH. PO7 Human Pathogenic Microsporidia: Detection and Genotyping in
HIV-positive and negative patients from Portugal. M. L. LOBO 1 , L.
XIAO2 , F. ANTUNES 3 , O. MATOS1 ; 1 Instituto de Higiene e
Medicina Tropical, CMDT, UNL, Lisboa, Portugal, 2 Centers for
Disease Control and Prevention, Atlanta, GA, USA, 3 Hospital de
Santa Maria, FM/UL, Lisboa, Portugal. PO8 QSAR Study of New
Inhibitors for Pneumocystis carinii Oxidosqualene Cyclase. ALEKSEY
POROLLO1 , MARGARET S. COLLINS1,2 , MELANIE T. CUSHION1,2 ; 1
University of Cincinnati College of Medicine Department of
Environmental Health, Cincinnati, OH, USA; 2 University of
Cincinnati Department of Internal Medicine and the Cincinnati
Veterans Affairs Medical Center, Cincinnati, OH PO9 Factors
Influencing the Carriage, Colonization, and Transmission of
Pneumocystis carinii. K.A. LYNCH, and M.T. CUSHION, University of
Cincinnati College of Medicine and the Cincinnati Veterans Affairs
Medical 6. 6 Center, Cincinnati, Ohio, USA. PO10 Microscopic
Studies of Anncaliia algerae-Infected Cell Cultures and Analysis of
its Beta-Tubulin Gene Suggest Albendazole Sensitivity. MARIANITA
SANTIANA, PETER TAKVORIAN, CYRILLA PAU, ANN CALI. Rutgers
University, Newark, NJ, USA. PO11 Cryptosporidium spp. in Pet Birds
in Henan, China: Prevalence and Molecular Characterizations. MENG
QI,1 RONGJUN WANG,1 CHANGSHEN NING,1 LONGXIAN ZHANG,1 FUCHUN JIAN,1
YANRU SUN,1 LIHUA XIAO2 ; 1 The College of Animal Science and
Veterinary Medicine, Henan Agricultural University, Zhengzhou
450002, China, 2 Division of Foodborne, Bacterial, and Mycotic
Diseases, National Center for Emerging and Zoonotic Infectious
Diseases, Centers for Disease Control and Prevention, Atlanta, GA
30333, USA PO12 Standardization of PCR Primers for gp60-based
Subtyping of Cryptosporidium hominis and Cryptosporidium parvum.
SATOMI KATO, LIHUA XIAO, Atlanta Research and Education Foundation,
Centers for Disease Control and Prevention. PO13 Preliminary
Biochemical Data on a Type II Thioesterase from Cryptosporidium
parvum (CpTEII). FENGGUANG GUO, GUAN ZHU; Department of Veterinary
Pathobiology, Texas A&M University, TX, USA. PO14
Hospital-Based Monitoring of Cryptosporidium parvum, Giardia
lamblia and Entamoeba histolytica in the Republic of Korea,
2004-2008. HYENG-IL CHEUN*, YI-YOUNG LIM, SHIN-HYEONG CHO, JUNG-WON
JU, SANG- EUN LEE, JEONG-YEON KIM, WON-JA LEE. Division of Malaria
and Parasitic Diseases, National Institute of Health, Korea Centers
for Disease Control and Prevention, Seoul 122-701, Korea. PO15 Low
Prevalence of Pneumocystis Pneumonia (PCP) but High Prevalence of
Pneumocystis Dihydropteroate Synthase (DHPS) Gene Mutations in
HIV-Infected Persons in Uganda. LAURENCE HUANG,1 STEVE M. TAYLOR,2
WILLIAM WORODRIA,3 , LEAH JARLSBERG,1 J. LUCIAN DAVIS,1 ADITHYA
CATTAMANCHI,1 SAMUEL D. YOO,3 ALFRED ANDAMA,3 SASKIA DEN BOON,3
RACHEL KYEYUNE,3 STEVEN MESHNICK,2 . 1 San Francisco, CA, USA, 2
Chapel Hill, NC, USA, 3 Kampala, Uganda. PO16 Variation in the
erg11 Gene from Pneumocystis jirovecii. SCOTT P. KEELY,1 JAMES R.
STRINGER,1 CARLO CONTINI,2 BETTINA LUNGREN,3 RONALD BRUBAKER4 ,
LAURA Q. JOHNSTON,5 EDNA S. KANESHIRO5 ; 1 Dept. Molecular
Genetics, Biochemistry & Microbiology, 5 Dept. Biological
Sciences, Univ. Cincinnati, Cincinnati, OH; 2 Dept. Clinical &
Experimental Medicine, Univ. Ferrara, Ferrara, Italy, 3 Dept.
Clinical Microbiology, Hvidovre Univ. Hospital, Hvidovre, Denmark.
4 Department of Pathology, Christ Hospital, Cincinnati, OH. 7. 7
PO17 Development of a Multilocus Sequence Typing Tool for
Cryptosporidium muris and Cryptosporidium andersoni. YAOYU FENG,1
WENLI YANG,2 UNA RYAN,3 LONGXIAN ZHANG,4 MARTIN KV,5 BETISLAV
KOUDELA,5 NA LI,2,6 RONALD FAYER,7 LIHUA XIAO2 ; 1 East China
University of Science and Technology, Shanghai, China, 2 Centers
for Disease Control and Prevention, Atlanta, GA, USA, 3 Murdoch,
Western Australia, Australia, 4 Henan Agricultural University,
Zhengzhou, China, 5 Academy of Sciences of the Czech Republic, esk
Budjovice, Czech Republic, 6 Tongji University, Shanghai, China,7
U.S. Department of Agriculture, Beltsville, Maryland, USA. PO18
Amoebicidal and Amebastatic Activity In Vitro of the Resin of
Gymnosperma glutinosum Against Acanthamoeba castellanii
trophozoites. RODRIGUEZ- MONROY MA1 , PEA-JUAREZ MC2 , OMAA-MOLINA
M1 , GONZALEZ- ROBLES A3 , SALAZAR-VILLATORO L3 , CANALES-MARTINEZ
MM4 .1 Profesor Carrera de Medicina, U.N.A.M. F.E.S. Iztacala. 2
Profesor Carrera de Biologa, U.N.A.M. F.E.S. Iztacala. 3
Departamento de Infectmica y Patognesis Molecular, CINVESTAV,
I.P.N., 4 Profesor Carrera de Medicina, U.N.A.M. F.E.S. Iztacala,
Edo. Mexico. 12:00 NOON LUNCH QUEEN'S COURT PM 1:00 SYMPOSIUM "HOST
RESPONSES TO IMMUNODEFICIENCY-ASSOCIATED DISEASE-CAUSING PROTISTS"
MOKU OLA I 1:00 S1 Introduction CHAO-HUNG LEE, Department of
Pathology & Laboratory Medicine, Indiana University School of
Medicine, Indianapolis, IN, USA. 1:30 S2 Role of Host and Parasite
Glycans and Glycan-Binding Proteins in Cryptosporidium-Host Cell
Interactions. HONORINE D. WARD, NAJMA BHAT, ROBERTA M. CO'CONNOR,
Division of Geographic Medicine and Infectious Diseases, Tufts
Medical Center, Tufts University School of Medicine, Boston, MA
02111. 2:15 S3 Host Defense against Pneumocystis in Neonates:
Lessons for Adults. BETH A. GARVY; Departments of Microbiology,
Immunology, and Molecular Genetics and Internal Medicine Division
of Infectious Diseases, University of Kentucky and VA Medical
Center, Lexington, KY. 3:00 BREAK 8. 8 3:30 S4 Plasmacytoid
Dendritic Cells in Aging Animals Down-Regulate Conventional
Dendritic Cell Response against Encephalitozoon cuniculi Infection.
JASON P GIGLEY, MAAZ SOHAIL, IMTIAZ A KHAN; Department of
Microbiology, Immunology and Tropical Medicine George Washington
University Medical Center, Washington DC. 4:15 S5 Toxoplasma gondii
Infection and Neuropsychiatric Disease. CRAIG W. ROBERTS;
Biomedical Sciences, University of Strathclyde, Glasgow, UK. 5:00
REMOVE POSTER SESSION A POSTERS 5:30 SET UP POSTER SESSION B
POSTERS EVENING ORGANISM-BASED DINNERS TUESDAY, AUGUST 3, 2010 AM
7:30 REGISTRATION FOYER, LOWER LEVEL PLATFORM SESSION II MOKU OLA I
CO-CHAIRS: MALCOLM FINKELMAN, BETTINA LUNDGREN 8:00 PL7 Development
of a Multilocus Sequence Typing Tool for High Resolution Genotyping
of Enterocytozoon bieneusi. YAOYU FENG,1 NA LI,2,3 THERESA DEAREN,
MARIA LUSA LOBO4 , OLGA MATOS4 , 2 LIHUA XIAO3* ; 1 East China
University of Science and Technology, Shanghai, China, 2 Centers
for Disease Control and Prevention, Atlanta, GA, USA, 3 Tongji
University, Shanghai, China, 1 Instituto de Higiene e Medicina
Tropical, CMDT, UNL, Lisboa, Portugal. 8:20 PL8 Identification of
Proteins of the Microsporidian Invasion Apparatus. KAYA GHOSH,1 ,
ANN CALI,2 PETER M. TAKVORIAN,2 RUTH HOGUE- ANGELLETI,3 LOUIS M.
WEISS1,4 . Departments of 1 Pathology, 3 Developmental and
Molecular Biology, and 4 Medicine, Albert Einstein College of
Medicine, Bronx, NY 10461, 2 Department of Biological Sciences,
Rutgers University, Newark, NJ 07102. 8:40 PL9 Molecular Genotyping
of Viable Waterborne Protozoa. CRISTIN C. BRESCIA1 , UKO NWA3 ,
SHANNON M. GRIFFIN1 , MICHAEL W. WARE1 , EUNICE A.VARUGHESE1 ,
ANDREY I. EGOROV2 , ERIC N. VILLEGAS1,3 , 1 National Exposure
Research Laboratory, 2 National Center for Environmental
Assessment, US Environmental Protection Agency, Cincinnati, OH
45268, 3 Department of Biological Sciences, University of
Cincinnati, Ohio 45221. 9. 9 9:00 PL10 Evolutionary Affiliation,
but Metabolic Diversity between Cryptosporidium and Ascogregarina
taiwanensis. GUAN ZHU,1 THOMAS J. TEMPLETON,2 SHINICHIRO ENOMOTO,3
MITCHELL S. ABRAHAMSEN,3 WEI-JUNE CHEN4 ; 1 Department of
Veterinary Pathobiology, College of Veterinary Medicine &
Biomedical Sciences, Texas A&M University, College Station; TX,
USA, 2 Department of Microbiology and Immunology, Weill Cornell
Medical College, and the Program in Immunology and Microbial
Pathogenesis, Weill Graduate School of Medical Sciences of Cornell
University, New York, NY, USA, 3 Department of Veterinary &
Biomedical Sciences, University of Minnesota, St. Paul, MN, USA, 4
Department of Public Health and Parasitology, College of Medicine,
Chang Gung University, Kwei-San, Tao- Yuan, Taiwan. 9:20 PL11
Metabolic Targets for Controlling Pneumocystis Adenosylmethionine
Supply. SALIM MERALI, Department of Biochemistry, Temple University
School of Medicine, Philadelphia, Pennsylvania 19140. 9:40 PL12
Pneumocystis jirovecii Multilocus Genotyping using High Throughput
Methodologies for Large Scale SNPs Screening. F. ESTEVES1 , J.
GASPAR2 , F. ANTUNES3 , K. MANSINHO4 and O. MATOS1 ; 1 Unidade de
Protozorios Oportunistas/VIH e Outras Protozooses CMDT, Instituto
de Higiene e Medicina Tropical, Universidade Nova de Lisboa, 2
Departamento de Gentica, Faculdade de Cincias Mdicas, Universidade
Nova de Lisboa, 3 Clinica Universitria de Doenas Infecciosas,
Faculdade de Medicina, Universidade de Lisboa, Hospital de Santa
Maria, 4 Servio de Doenas Infecciosas, Hospital de Egas Moniz,
Lisboa, Portugal. 10:00 FIELD TRIPS/TOURS/FREE TIME AKAKA FALLS
& BOTANICAL GARDENS FIELD TRIP (w/ Lunch) HAWAII VOLCANO
NATIONAL PARK FIELD TRIP (w/ Lunch) EVENING ORGANISM-BASED DINNERS
WEDNESDAY, AUGUST 4, 2010 AM 7:30 REGISTRATION FOYER, LOWER LEVEL
PLATFORM SESSION III MOKU OLA I CO-CHAIRS: ANN CALI, SALIM MERALI
8:00 PL13 Pseudoloma neurophilia and Pleistophora hyphessobriconis;
the Causes of Two Important Microsporidial Diseases in Zebrafish,
Danio rerio. MICHAEL L. KENT, JUSTIN SANDERS, VIRGINIA WATRAL,
Department of Microbiology, Oregon State University, Corvallis,
Oregon. 10. 10 8:20 PL14 HIV Treatment as Prevention for the
Development of Protist Infections: Year 2010. JENS LUNDGREN,
University of Copenhagen and State University Hospital, Panum
Institute, 2200 Copenhagen N, Denmark. 8:40 PL15 An
Adhesion-Induced Signaling Cascade in Pneumocystis carinii:
Potential Role of the Downstream PcAce2 Transcription Factor in
Cell Wall Remodeling. ANDREW H. LIMPER, THEODORE J. KOTTOM;
Thoracic Diseases Research Unit, Mayo Clinic, Rochester, MN, USA.
9:00 PL16 Heterologous Systems Expressing the Recombinant
Pneumocystis carinii S- Adenosyl-L-Methionine: Sterol C-24
Methyltransferase. LAURA Q. JOHNSTON, EDNA S. KANESHIRO, Dept.
Biological Sciences, Univ. Cincinnati, Cincinnati, OH, USA. 9:20
PL17 Sequences of the erg11 Gene from P. jirovecii, P. carinii, P.
murina and Pneumocystis from the Rhesus Macaque Monkey. SCOTT
KEELY,1 JAMES R. STRINGER,1 MICHAEL J. LINKE,3 TAKAHISA FURUTA,4
EDNA S. KANESHIRO2 . 1 Dept. Molecular Genetics, Biochemistry &
Microbiology, and 2 Dept. Biological Sciences, Univ. Cincinnati,
Cincinnati, OH, 3 Veterans Affairs Medical Center, Cincinnati, OH,
4 Dept. Microbiology and Immunology, Institute of Medical Science,
Univ. Tokyo, Tokyo, Japan. 9:40 PL18 Molecular Epidemiology of
Cryptosporidium and Giardia in Children in Shanghai, China. FENG,
Y.Y.1* , WANG, L2 , DUAN, L.P.2 , GOMEZ-PUERTA, L.A.2 , XIAO, L.2
*, 1 East China University of Science and Technology, Shanghai
200237, China; 2 Centers for Disease Control and Prevention,
Atlanta, Georgia 30341, USA. 10:00 BREAK 10:30 POSTER SESSION B
#19-36 PO19 DNA Replication Continues in Pneumocystis carinii
Trophic Forms Treated with the -1,3-Glucan Synthesis Inhibitor
Pneumocandin L-693,989. MICHAEL A. WYDER, LAURA Q. JOHNSTON, EDNA
S. KANESHIRO, Department of Biological Sciences, University of
Cincinnati, Cincinnati, OH, USA. PO20 Pneumocystis carinii Cell
Wall Chitins Induce Host Lung Responses. L.R. VILLEGAS, T.J.
KOTTOM, A.H. LIMPER, Thoracic Disease Research Unit, Mayo Clinic
College of Medicine, Rochester, MN 55905. PO21 Alternative
Activation of Alveolar Macrophages and Alteration of NFB Signaling
Results in a Reduced Response to Pneumocystis Infection by
Neonates. CATE KURKJIAN,1,2 MELISSA HOLLIFIELD,1,2 BRIAN S.
MURPHY,1 BETH A. GARVY,1,2 . 1 Department of Microbiology,
Immunology and Molecular 11. 11 Genetics and Division of Infectious
Diseases, University of Kentucky, and 2 VA Medical Center,
Lexington, KY. PO22 Radiation-Induced Alteration of the
Cryptosporidium parvum Oocyst Proteome. SOO-UNG LEE1 , MIKYO JOUNG1
, KYOUNGJIN CHOI1 , WOO-YOON PARK2 , YOUNG-HOON JI3 , JAE-RAN YU1 .
1 Konkuk University, School of Medicine Seoul, Republic of Korea, 2
College of Medicine, Chungbuk National University, Cheongju,
Republic of Korea, 3 Korea Institute of Radiological and Medical
Sciences, Seoul, Republic of Korea PO23 An Approach to Investigate
the Proteome of Pneumocystis carinii. VICENTE FRIAZA1 , ANNA
MARTINEZ2,3 , CARMEN DE LA HORRA1 , CECILE- MARIE ALIOUAT-DENIS2,3
, NIEVES RESPALDIZA1 , ANNIE VITSE2,3 , RUBEN MORILLA1 , ELENA
CAMPANO1 , EDUARDO DEI-CAS2,4 , ENRIQUE J. CALDERON1 ; 1 Instituto
de Biomedicina de Sevilla and CIBER de Epidemiologia y Salud
Pblica. Virgen del Rocio University Hospital, Seville, Spain, 2
Biology and Diversity of Emergent Eukaryotic Pathogens (BDEEP)
(EA3609), IFR142, Institut Pasteur de Lille & 3 Department of
Parasitology- Mycology, Faculty of Biological and Pharmaceutical
Sciences, University of Lille Nord de France, Lille, France, 4
Department of Parasitology-Mycology, Faculty of Medicine,
University of Lille Nord de France, Biology-Pathology Centre,
University Hospital Center, Lille, France. PO24 Comparison of
Proteomic Profiles in the Bronchoalveolar Lavage Fluid of
Idiopathic Pulmonary Fibrosis Patients with and without
Pneumocystis Colonization. VICENTE FRIAZA1 , CARMEN DE LA HORRA1 ,
RUBEN MORILLA1 , NIEVES RESPALDIZA1 , MARCO A. MONTES-CANO1 , LAURA
RIVERO1 , SONIA GUTIERREZ1 , ELENA CAMPANO1 , ISABEL MARTIN-
GARRIDO1 , JOSE M.VARELA1 , FRANCISCO J. MEDRANO1 , JOSE
MARTIN-JUAN1 , JUAN PARRADO2 , JUAN BAUTISTA2 , ENRIQUE J.
CALDERON1 ; 1 Instituto de Biomedicina de Sevilla and CIBER de
Epidemiologia y Salud Pblica. Virgen del Rocio University Hospital,
Seville, Spain. 2 Departamento de Bioqumica y Biologa Molecular,
Universidad de Sevilla, Seville, Spain. PO25 Association of
Idiopathic Pulmonary Fibrosis Severity and Pneumocystis jirovecii
Colonization. NIEVES RESPALDIZA1 , ISABEL MARTIN-GARRIDO1 , EDUARDO
MARQUEZ-MARTIN2 , VICENTE FRIAZA1 , SONIA GUTIERREZ1 , RUBEN
MORILLA1 , LAURA RIVERO1 , ELENA CAMPANO1 , RAFAEL TERAN1 , MARCO
A. MONTES-CANO1 , JOSE M.VARELA1 , FRANCISCO J. MEDRANO1 , JOSE
MARTIN-JUAN2 , CARMEN DE LA HORRA1 , ENRIQUE J. CALDERON1 ; 1
Instituto de Biomedicina de Sevilla and CIBER de Epidemiologia y
Salud Pblic; 2 Servicio de Neumologa, Virgen del Rocio University
Hospital, Seville, Spain. PO26 Treatment of Pneumocystis carinii
with Sodium Nitrite in Suspension or Biofilm Cultures Dramatically
Reduces Viability. M.T. CUSHION1,2 , M.S. COLLINS1,2 , 12. 12 D.J.
HASSETT3 ; 1 University of Cincinnati College of Medicine,
Department of Internal Medicine; 2 Cincinnati Veterans Affairs
Medical Center, Cincinnati, Ohio; 3 University of Cincinnati
College of Medicine, Department of Molecular Genetics, Biochemistry
and Microbiology, Cincinnati, OH PO27 The Effect of Oxygen on
Viability, Sterol Uptake, and Transcriptional Responses in
Pneumocystis carinii. TIFFANY M. JOFFRION,1,* MARGARET S. COLLINS,1
MELANIE T. CUSHION1,2 . 1 University of Cincinnati College of
Medicine, Cincinnati, OH 45267, USA; 2 Veterans Administration
Medical Center, Cincinnati, OH 45220, USA. PO28 Morphometrics of
Assemblages of Giardia duodenalis Cysts from the Feces of Dogs and
Cats. HEEJEONG YOUN,1 DWIGHT D. BOWMAN,2 STEPHANIE B. YAGER,2
BRITTA A. OKYERE,2 BO LI,2 DAVID KYUHYUNG KANG,2 MARISSA KARPOFF,2
HYUN JI KIM,2 HUSSNI O. MOHAMMED,2 ARACELI LUCIO-FORSTER,2 JANICE
L. LIOTTA.2 1 College of Veterinary Medicine, Seoul National
University, Seoul, Korea; 2 Department of Microbiology &
Immunology, College of Veterinary Medicine, Cornell University,
Ithaca, NY, USA. PO29 Use of the Luna Stain Allows Specific, Rapid
and Unequivocal Detection of Pseudoloma neurophilia Spores in
Formalin Fixed Zebrafish Tissue Sections. TRACY S. PETERSON,
MICHAEL L. KENT, Department of Microbiology, Oregon State
University, Corvallis, Oregon. PO30 Seroprevalence and in vitro
Isolation of Toxoplasma gondii from Chronically Infected Pigeons in
Portugal. HELGA WAAP1 , RITA CARDOSO2,3 , HELENA NGELO4 , ANABELA
VILARES4 , HELDER CORTES5 , JOS MEIRELES3 , ALEXANDRE LEITO2 ; 1
Instituto Nacional de Recursos Biolgicos-LNIV, Lisbon, Portugal, 2
Instituto de Investigao Cientfica e Tropical, CIISA, Lisbon,
Portugal, 3 Faculdade de Medicina Veterinria, CIISA, Lisbon,
Portugal, 4 Instituto Nacional de Sade Dr. Ricardo Jorge, Lisbon,
Portugal, 5 Laboratrio de Parasitologia Victor Caeiro (ICAAM)
Universidade de vora, Portugal. PO31 Prevalence, Genetic
Characteristics, and Zoonotic Potential of Cryptosporidium in Farm
Rabbits in China. KE SHI1 , FUCHUAN JIAN1 , CHAOCHAO LV1 ,
CHANGSHEN NING1 , LONGXIAN ZHANG1, 2, 3 *, XUPENG REN1 , THERESA K.
DEAREN2 , NA LI2 , MENG QI1 , LIHUA XIAO2 *; 1 College of Animal
Science and Veterinary Medicine, Henan Agricultural University,
Zhengzhou 450002, P. R. China, 2 Atlanta Research and Education
Foundation, 1670 Clairmont Road, Decatur, GA 30033, USA, 3 Division
of Foodborne, Bacterial and Mycotic Diseases, National Center for
Emerging and Zoonotic Infectious Diseases, Centers for Disease
Control and Prevention, Atlanta, GA 30333, USA. PO32
Cryptosporidium parvum Induces an Invasive Intestinal
Adenocarcinoma in 13. 13 Immunosupressed Hosts. GABRIELA CERTAD,1
SADIA BENAMROUZ- HAMRAOUI,1, 2 KARINE GUYOT,1 ANTHONY MOURAY,3
THIERRY CHASSAT,3 BAPTISTE DELAIRE,4 VALERIE CONSEIL,2 EDUARDO DEI-
CAS,1, 5 COLETTE CREUSY,4 ; 1 BDPEE-EA3609- Biologie et Diversit
des Pathognes Eucaryotes Emergents, IFR142-Institut Pasteur de
Lille France, 2 Laboratoire Environnement et Sant, FLST, Universit
Catholique de Lille, Universit Lille Nord-de-France, 3 Plateau
dExprimentation Animale, Institut Pasteur de Lille, France, 4
Service dAnatomie et Cytologie Pathologiques, Groupe Hospitalier de
lUniversit Catholique de Lille, France, 5 Service dParasitologie-
Mycologie, CHRU de Lille, Universit Lille Nord-de-France, France.
PO33 Distribution of Pneumocystis Colonization in the Human Lung.
SHEILA SIVAM,1 FRANK C. SCIURBA,1 LORRIE LUCHT,1 YINGZE ZHANG,1
STEVEN R. DUNCAN,1 KAREN A NORRIS,2 ALISON MORRIS1,2 . 1 Department
of Medicine, 2 Department. of Immunology, University of Pittsburgh,
Pittsburgh, Pennsylvania. PO34 Isolation and Biological
Characterization of Free-Living Amoeba Isolated from Clinical Cases
of Amoebic Keratitis, Contact Lenses and Lens Solutions. MARITZA
OMAA-MOLINA1 , REN MNDEZ-CRUZ1 , ARTURO GONZLEZ-ROBLES2 , MARCO
RODRGUEZ-MONROY1 , ELIZABETH RAMREZ-FLORES1 , MA. DOLORES
HERNNDEZ-MARTNEZ1 , MA. ESTHER INIESTRA-SOLRZANO1 , ALEXANDER
BERNAL-ESCOBAR1 . 1 School of Superior Studies Iztacala, UNAM. Los
Reyes Iztacala, Tlalnepantla, Mexico State, Mexico. 2 Department of
Infectomic and Molecular Pathogenesis Center for Research and
Advanced Studies, Mexico City, Mexico. PO35 Pneumocystis jirovecii
Colonization in Elderly Adults Admitted to the Hospital with
Community-Acquired Pneumonia in Santiago, Chile. MARILZ HERNANDEZ,
PATRICIA PIZARRO, ANDREA ARAYA, REBECA BUSTAMANTE, CAROLINA A.
PONCE, SERGIO L. VARGAS. University of Chile School of Medicine,
Santiago, Chile. PO36 Molecular Epidemiology of Cryptosporidium in
HIV Patients in Henan, China. LING WANG,1,2 XUDONG ZHAO,3 HONGWEI
ZHANG,3 LONGXIAN ZHANG,4 LIHUA XIAO,2 YAOYU FENG1 ; 1 East China
University of Science and Technology, Shanghai, China, 2 Centers
for Disease Control and Prevention, Atlanta, Georgia, USA, 3 Henan
Provincial Center for Disease Control and Prevention, Zhengzhou,
China, 4 Henan Agricultural University, Zhengzhou, China. 12:00
LUNCH QUEEN'S COURT 14. 14 PM PLATFORM SESSION IV MOKU OLA I
CO-CHAIRS: SATOMI KATO, EVA CARMONA 1:00 PL19 Development of a
Multiplex-PCR/Single Base Extension Methodology for Pneumocystis
jirovecii Genotyping. F. ESTEVES1 , J. GASPAR2 , F. ANTUNES3 , K.
MANSINHO4 and O. MATOS1 ; 1 Unidade de Protozorios Oportunistas/VIH
e Outras Protozooses CMDT, Instituto de Higiene e Medicina
Tropical, Universidade Nova de Lisboa, 2 Departamento de Gentica,
Faculdade de Cincias Mdicas, Universidade Nova de Lisboa, 3 Clinica
Universitria de Doenas Infecciosas, Faculdade de Medicina,
Universidade de Lisboa, Hospital de Santa Maria, 4 Servio de Doenas
Infecciosas, Hospital de Egas Moniz, Lisboa, Portugal. 1:20 PL20
Propylene Glycol Induces Pseudocyst Formation in Acanthamoeba spp.
JARMILA KLIESCIKOVA, EVA HORACKOVA, EVA NOHYNKOVA, Department of
Tropical Medicine, 1st Medical Faculty, Charles University in
Prague and Faculty Hospital Bulovka, 128 00 Prague 2, Czech
Republic. 1:40 PL21 Autophagy-Related Processes in Toxoplasma
gondii. DEBASISH GHOSH, JULIA WALTON, ANTHONY P. SINAI*. Department
of Microbiology, Immunology and Molecular Genetics, University of
Kentucky College of Medicine, 800 Rose St. Lexington, KY 40536.
2:00 ROUND TABLE DISCUSSION I "FUTURE OF FUNDING FOR OPPORTUNISTIC
PROTIST RESEARCH" MOKU OLA I CO-CHAIRS: ANTHONY SINAI; MELANIE T.
CUSHION 3:00 BREAK PLATFORM SESSION V MOKU OLA I CO-CHAIRS: NICOLE
MEISSNER, SCOTT KEELY 3:30 PL22 Evaluating the Effects of Chemical
Sanitizers and UV Irradiation on Toxoplasma gondii Oocyst
Viability. MICHAEL W. WARE,1 SWINBURNE A. J. AUGUSTINE,1 DAVID O.
ERISMAN,1 LEAH FOHL VILLEGAS,1 MARY JEAN SEE,2 SAMUEL L. HAYES,3
LARRY WYMER,1 H. D. ALAN LINDQUIST,4 FRANK W. SCHAEFER III,4 J. P.
DUBEY,5 ERIC N. VILLEGAS1,2* ; 1 National Exposure Research
Laboratory, 3 National Risk Management Research Laboratory, 4
National Homeland Security Research Center, U.S. Environmental
Protection Agency, Cincinnati, OH 45268; 2 Department of Biological
Sciences, University of Cincinnati, Cincinnati, OH 15. 15 45220; 5
Animal Parasitic Disease Laboratory, Agricultural Research Service,
U.S. Department of Agriculture, Beltsville, MD, 20705 3:50 PL23
Identification of the Microsporidium Anncaliia algerae as the
Causative Agent of a Human Vocal Cord Infection. ANN CALI,1 RONALD
NEAFIE,2 LOUIS M. WEISS,3 KAYA GHOSH,4 RACHNA GUPTA,5 REBECCA B.
VERGARA,6 PETER M. TAKVORIAN,1,3 . 1 Department of Biological
Sciences, Rutgers University, Newark, NJ 07102, 2 American
International Pathology Laboratories, Silver Spring, MD 20910, 3
Department of Pathology, Division of Parasitology and Tropical
Medicine, Albert Einstein College of Medicine, Bronx, NY 10461, 4
Department of Microbiology and Immunolgy, Albert Einstein College
of Medicine, Bronx, NY 10461, 5 ID Care, 105 Raider Blvd. Suite
101, Hillsborough, NJ 08844, 6 Department of Pathology, Newton
Memorial Hospital, Newton NJ 07860 . 4:10 PL24 Wildtype CD4 T Cells
have Increased Bcl2 Expression in Response to Pneumocystis
Infection. MICHAEL M. OPATA, MELISSA L. HOLLIFIELD, BETH A. GARVY;
Department of Microbiology, Immunology and Molecular Genetics,
University of Kentucky College of Medicine and VA Medical Center,
Lexington, KY 40536 USA. 4:30 PL25 Diagnosis and Age Distribution
of Primary Infection by Pneumocystis jirovecii: A Study on
Autopsied Lungs. SERGIO L. VARGAS1 ; CAROLINA .A. PONCE1 ; J.
ASTORGA3, M. GALLO2 ; MAGALI CHABE3 ; ISABELLE DURAND- JOLY3 , EL
MOUKHTAR ALIOUAT3 , EDUARDO DEI CAS3 . 1 Biomedical Sciences
Institute, University of Chile School of Medicine, 2 Servicio Mdico
Legal, Santiago, Chile and, 3 Pasteur Institute of Lille, France.
4:50 PL26 Genetic Mapping of Virulence Genes of Cryptosporidium
hominis. NA LI,1,2,3 LIHUA XIAO,1 VITALIANO A. CAMA,1 YNES ORTEGA,4
ROBERT H. GILMAN,5 YAOYU FENG6 ; 1 Centers for Disease Control and
Prevention, Atlanta, GA, USA, 2 Atlanta Research and Education
Foundation, Atlanta, GA, USA, 3 Tongji University, Shanghai, China,
4 University of Georgia, Griffin, GA, USA, 5 Johns Hopkins
University, Baltimore, MD, USA, 6 East China University of Science
and Technology, Shanghai, China. 6:00 INTERNATIONAL DINNER W/
ENTERTAINMENT MOKU OLA BALLROOM AND GARDENS THURSDAY, AUGUST 5,
2010 AM 7:30 REGISTRATION FOYER, LOWER LEVEL 16. 16 PLATFORM
SESSION VI MOKU OLA I CO-CHAIRS: SHEILA SIVAM, GUAN ZHU 8:00 PL27
Pneumocystis (13)--D-Glucan (BG): Role in Diagnosis and Potential
Contribution to Pathophysiology. MALCOLM A. FINKELMAN; Associates
of Cape Cod, Inc., Falmouth, MA, USA 8:20 PL28 The Biochemistry and
Genetics of Mouse Resistance to Toxoplasma gondii: the Role of
Immunity-Related GTPases (IRG proteins). J. P. HUNN, A. KHAMINETS,
J. LILUE, B. MUELLER, N. PAPIC, N. PAWLOWSKI, T. STEINFELDT, J. C.
HOWARD; Institute for Genetics, University of Cologne, Cologne,
NRW, Germany. 8:40 PL29 Acanthamoeba culbertsoni Elicits Soluble
Factors that Exert Anti-Microglial Cell Activity. FRANCINE
MARCIANO-CABRAL, JENICA L. HARRISON, GABRIELA A. FERREIRA, ERINN S.
RABORN, AUDREY D. LAFRENAYE, GUY A. CABRAL; Virginia Commonwealth
University, School of Medicine, Richmond, VA, USA 9:00 PL30
Functional Complementation of Saccharomyces cerevisiae erg11
Deletion Mutant by the Pneumocystis carinii erg11 cDNA and
Comprehensive Identification of >20 Sterols in Wild Type and
Transformed Yeast Cells. STEPHENSON W. NKININ,1 JAMES R. STRINGER,2
SCOTT KEELY,2 KENNETH D.R. SETCHELL,3 JOS-LUIS GINER,4 EDNA S.
KANESHIRO1 . 1 Dept. Biological Sciences, and 2 Dept. Molecular
Genetics, Biochemistry & Microbiology, University of
Cincinnati, Cincinnati, OH; 3 Dept. Pediatrics, Cincinnati
Children's Hospital Medical Center, Cincinnati, OH, 4 Dept.
Chemistry, SUNY-ESF, Syracuse, NY, USA. 9:20 PL31 Effect of
Calmodulin on Phagocytic Activity of Alveolar Macrophages during
Pneumocystis Pneumonia. CHUNG-PING LIAO, JINGHONG WANG, PAMELA J.
DURANT, CHAO-HUNG LEE,* Department of Pathology and Laboratory
Medicine, Indiana University School of Medicine, Indianapolis,
Indiana 46202, USA 9:40 BREAK PLATFORM SESSION VII MOKU OLA I
CO-CHAIRS: LAURENCE HUANG, TIFFANY JOFFRION 10:10 PL32
MicrosporidiaDB a New Integrated Genomic Resource. LOUIS M.
WEISS,1,2 OMAR S. HARB,3 DAVID S. ROOS3 for the EuPathDB Team 3,4 .
Departments of 1 Pathology and 2 Medicine, Albert Einstein College
of Medicine, Bronx, NY, 10461; 3 Department of Biology, University
of Pennsylvania, Philadelphia, PA, 17. 17 19104., 4 Department of
Genetics, University of Georgia, Athens, GA, 30602. 10:30 PL33
Adherence and Internalization of Cryptosporidium parvum Oocysts
Associated with Fresh Produce. RONALD FAYER, DUMITRU MACARICIN,
GARY BAUCHAN, MONICA SANTIN-DURAN; USDA, Agricultural Research
Service, Beltsville, Maryland, USA. 10:50 PL34 Blastocystis hominis
Induces Increased Permeability of HT-29 Polarized Monolayers and
Resists Phagocytosis by Macrophages. LUIZ BERMUDEZ,1,2 VIRGINIA
WATRAL,2 BRITTANY MORGAN,2 SASHA ROSE,1 MICHAEL KENT 2,1 , 1
Department of Biomedical Sciences, College of Veterinary Medicine,
Oregon State University, Corvallis OR, 97331, 2 Department of
Microbiology, Oregon State University, Corvallis OR, 97331 11:10
ROUND TABLE DISCUSSION II "NOMENCLATURE: HOW SHOULD WE ABBREVIATE
GENES, CDNA, GENE PRODUCTS, ETC. IN OPPORTUNISTIC PROTIST
PUBLICATIONS?. WHAT ARE BEING USED AND CAN WE ACHIEVE UNIFORMITY?"
CHAIR: ANDREW LIMPER, LOUIS WEISS DISCUSSANTS: MELANIE T. CUSHION,
LIHUA XIAO 12:00 NOON LUNCH QUEEN'S COURT PM PLATFORM SESSION VIII
MOKU OLA 1 CO-CHAIRS: YAOYU FENG, ERIC VILLEGAS 1:00 PL35
-Microsporidia Spore Adherence in a Mouse Animal Model. CORY A.
LEONARD, J. RUSSELL HAYMAN; East Tennessee State University, James
H. Quilllen College of Medicine, Johnson City, TN. 1:20 PL36
Transcriptomic Approaches to Study Stage-Specific Genes of
Pneumocystis carinii. ANNA MARTINEZ,1 MAGALI CHABE,1 MELANIE
CUSHION,2 CHRISTINE HUBANS,3 DAVID HOT,4 EDUARDO DEI-CAS,5 EL
MOUKHTAR ALIOUAT,1 CECILE-MARIE ALIOUAT-DENIS,1 ; 1 BDEEP-
EA3609-Laboratoire de Parasitologie-Mycologie, Facult de Pharmacie
de Lille, Universit Lille Nord-de-France & IFR142-Institut
Pasteur de Lille, France, 2 University of Cincinnati College of
Medicine, Dept of Internal Medicine, Division of Infectious
Diseases, Cincinnati Veterans Administration Medical Center,
Cincinnati, Ohio, USA, 3 Genoscreen Company, Campus de lInstitut
Pasteur de Lille, France, 4 TAG Transcriptomics and Applied
Genomics 18. 18 UMR8161, Institut Pasteur de Lille, France, 5
BDEEP-EA3609-IFR142-Institut Pasteur de Lille & Service de
Parasitologie-Mycologie, CHRU de Lille, Universit Lille
Nord-de-France, France. 1:40 PL37 Pneumocystis S-Adenosylmethionine
Transport. OSCAR PEREZ-LEAL,1 CAMILO MONCADA,1 ALLEN B. CLARKSON,2
SALIM MERALI1 , 1 Department of Biochemistry, Temple University
School of Medicine, Philadelphia, Pennsylvania 19140 and 2
Department of Medical Parasitology, New York University School of
Medicine, New York, New York 10016. 2:00 PL38 Pneumocystis
Occurrence in Wild Rodents from South East Asia. MAGALI CHABE,1
VINCENT HERBRETEAU,2 JEAN-PIERRE HUGOT,3 YANNICK CHAVAL,4 EDUARDO
DEI-CAS,5 SERGE MORAND6 ; 1 BDEEP-EA3609- Laboratoire de
Parasitologie-Mycologie, Facult de Pharmacie de Lille, Universit
Lille Nord-de-France & IFR142-Institut Pasteur de Lille,
France, 2 Cemagref, UMR TETIS, Montpellier, France & IRD,
UR178, Mahidol University, Nakhon Pathom, Thailand, 3 OSEB, UMR
5202 CNRS, Musum National dHistoire Naturelle, Paris, France, 4
Centre de Biologie et de Gestion des Populations (CBGP), Campus
International de Baillarguet, Montferrier sur Lez, France, 5
BDEEP-EA3609-IFR142-Institut Pasteur de Lille & Service de
Parasitologie-Mycologie, CHRU de Lille, Universit Lille
Nord-de-France, France, 6 Institut des Sciences de l'Evolution
CNRS-UM2, Universit Montpellier 2, France. 2:20 PL39 Yeast Proteome
Substrate Microarrays Provide Novel Insights into Pneumocystis
carinii PcCbk1 and PcSte20 Signaling Biology. THEODORE J. KOTTOM1 ,
ANDREW H. LIMPER1 ; 1 Thoracic Diseases Research Unit, Mayo Clinic,
Rochester, MN, USA. 2:40 CLOSING COMMENTS Looking to the Future -
LOUIS WEISS 3:00 BREAK 3:00-4:30 POSTER SESSION B TAKE DOWN MOKU
OLA II 19. 19 ABSTRACTS PL1 The Biochemistry and Genetics of Mouse
Resistance to Toxoplasma gondii: the Role of Immunity-Related
GTPases (IRG proteins). J. P. HUNN, A. KHAMINETS, J. LILUE, B.
MUELLER, N. PAPIC, N. PAWLOWSKI, T.STEINFELDT, J. C. HOWARD;
Institute for Genetics, University of Cologne, Cologne, NRW,
Germany. Toxoplasma gondii strains differ greatly in their
virulence for mice. Relatively avirulent strains are confronted by
the interferon-g- inducible IRG proteins that accumulate on the
cytosolic face of the parasitophorous vacuole (1). These vesiculate
and ultimately rupture the vacuolar membrane, leading to the death
of the parasite (1, 2). This confrontation attenuates the infection
and enables the parasite to effect the bradyzoite transition and
encystment without prejudice to the host. Loss of IRG proteins by
genomic disruption disturbs this equilibrium and greatly enhances
the virulence of genetically avirulent strains (3). Virulent
strains, by contrast, resist the initial loading of IRG proteins
onto the parasitophorous vacuole and replicate uncontrollably (4,
5). The IRG proteins are large, self-activating GTPases with
dynamin- like features. Their function depends on a dynamic control
system, exercised by three regulatory members of the family acting
as guanine nucleotide dissociation inhibitors, that maintains the
effector IRG proteins in the inactive GDP-bound (6). This control
is released upon infection with avirulent strains by the formation
of the parasitophorous vacuole membrane (PVM) on which effector IRG
proteins rapidly accumulate in the GTP- bound active form (7). It
follows that IRG proteins should be the targets for virulence
factors derived from T. gondii, and we will present evidence that
virulent strains can inactivate IRG proteins. We will also put the
dynamic relationship between genetically variable T. gondii strains
and mice into an ecological context and show that there is a
corresponding polymorphic variation in the IRG resistance proteins,
enabling certain wild mouse IRG genotypes to resist virulent T.
gondii strains. We argue that resistance to T. gondii is largely
driving the recent evolution of the IRG system in the mouse (8).
References: (1) Martens et al, 2005. PLoS Pathogens 1(3):e24 (73);
(2) Zhao, YO et al, 2009. PLoS Pathog 5(2): e1000288; (3) Taylor,
GA et al, 2007 Microbes Infect. 9:1644; (4) Zhao, YO et al, 2009.
Memorias do Instituto Oswaldo Cruz. 104(2):234-40; (5) Khaminets, A
et al, 2010, Cellular Microbiology, Epub ahead of print, PMID:
20109161; (6) Hunn, JP et al, 2008, EMBO Journal, 27(19):2495-509;
(7) Papic, N et al, 2008 J. Biological Chemistry 283(46):32143- 51;
(8) Hunn, JP and Howard, JC, 2010 PLoS Pathogens, in press. PL2
Involvement of Integrin Alpha 2 (ITGA2) in Cryptosporidium parvum
Infection. HAILI ZHANG, FENGGUANG GUO, GUAN ZHU; Department of
Veterinary Pathobiology, Texas A&M University, TX, USA.
Cryptosporidium parvum interacts with host cell membrane during
invasion and development. However, very little is known on the host
cell membrane proteins that interact with C. parvum. We have
previously observed from our microarray and qRT-PCR date that gene
expression profiles of several extracellular membrane (ECM)
proteins 20. 20 changed upon C. parvum infection, such as some host
cell integrin subunits. For example, the expression of integrin A2
(ITGA2) was increased by ~40% upon C. parvum infection, implying
that integrins might be involved in interacting with parasite. To
further test the hypothesis, we generated several stable ITGA2
knockdown lines of HCT-8 cells using plasmid-based RNA interference
technology. We observed constant ~20% reduction of parasite
invasion in the ITGA2-knockdown cells in comparison with controls.
Furthermore, parasite invasion was also decreased by 20-30% in
HCT-8 cell treated with ITGA2-specific antibodies. These
observations suggest that human ITGA2, and probably other integrin
subunits might participate in interacting with C. parvum. PL3
Lactosylceramide Mediates Pneumocystis Beta-Glucan Activation of
the Human IL- 23-IL-17 Axis. EVA M CARMONA, ANDREW H LIMPER.
Thoracic Diseases Research Unit, Mayo Clinic, Rochester, MN, USA.
Respiratory failure in Pneumocystis pneumonia is believed to be due
largely to an exaggerated host inflammatory response to the
organism rather than to the organism burden itself. Prior data from
our laboratory demonstrated that dendritic cells (DC) play an
important role orchestrating this inflammatory response. In
particular, stimulation of DC by Pneumocystis beta-glucans (PCBG)
induced a cytokine environment rich in IL-1 that favored a Th1
phenotype with subsequent IFN- production. Recent data have
revealed that fungal pathogens are also potent inducers of Th17
cells. Moreover, IL-17R deficient mice have poor defense against
some fungal infections. Therefore, this study was designed to
investigate whether PCBG-stimulated-DCs participate in Th17
differentiation and to study the molecular mechanisms that mediate
this process. In addition, we investigated the role of
lactosylceramide; a glycosphingolipid important in PC based
inflammation, in modulating this axis. Methods: Human DCs derived
from peripheral blood monocytes were stimulated with PCBG prior to
co- culture with nave CD4 cells. Cytokine secretion important for
Th17 differentiation was measured by ELISA in the supernatant of
these cells. NF-B activation was also analyzed in nuclear and
cytosolic extracts after stimulation with PCBG. A lactosylceramide
inhibitor, PDMP, was used to study the role of glycosphingolipids
as costimulatory molecule in PCBG induced IL- 23- IL7 axis.
Results: We observed that PCBG activates the IL-23-IL-17 axis.
Secondly, PCBG induces activation of both the classical and the
alternative pathways of NF- B in DC. We further demonstrated that
PDMP decreases IL-23 production through NF-B. Conclusions: These
findings provide evidence that PCBG matured-DC elicits activation
of the IL-23-IL-17 pathway though activation of both the classical
and the alternative pathways of NF-B. Our data also indicates that
lactosylceramide is crucial for activation of this pathway. Further
investigations are needed, but these studies provide further
evidence of lactosylceramide as new tool to modulate the
inflammatory response induced by PCBG in the immunocompromised
host. PL4 Pneumocystis carinii Lanosterol Synthase Requires
Saccharomyces cerevisiae 3- Ketoreductase for Localization to Lipid
Particles in Yeast. TIFFANY M. JOFFRION,1 MELANIE T. CUSHION1,2 . 1
University of Cincinnati College of Medicine, Cincinnati, OH 45267,
USA; 2 Veterans Administration Medical Center, Cincinnati, OH
45220, USA. Organisms in the genus Pneumocystis are ubiquitous,
opportunistic fungi capable of 21. 21 causing a lethal pneumonia in
immunocompromised mammalian hosts. Pneumocystis spp. are unique
members of the fungal kingdom due to the absence of ergosterol in
their cellular membranes. Although thought to obtain cholesterol by
scavenging, transcriptional analyses indicate that Pneumocystis
carinii encodes gene homologs involved in sterol biosynthesis. To
better understand the sterol pathway in these uncultivable fungi,
yeast deletion strains were used to interrogate the function and
localization of P. carinii lanosterol synthase (ERG7) to lipid
particles, which requires a 3- ketoreductase (ERG27) in yeast.
Expression of PcErg7p in an ERG7 null mutant of the yeast
Saccharomyces cerevisiae did not alter its growth rate and produced
functional lanosterol synthase, as evidenced by the presence of
lanosterol detected by gas chromatographic analysis in levels
comparable to that produced by the yeast enzyme. Western blotting
revealed that like the S. cerevisiae Erg7p, the PcErg7p localized
to lipid particles in yeast with a functional ERG27p, but this
trafficking was interrupted in an ERG27 deletion strain. In P.
carinii, the presence of PcErg7p in apparent lipid particles was
detected by fluorescence microscopy. By using the yeast
heterologous system, we show for the first time that the P. carinii
lanosterol synthase localizes to lipid particles and that this
targeting requires the 3-ketoreductase encoded by ERG27. Moreover,
the yeast heterologous system should be a useful tool for further
analysis of the P. carinii sterol pathway. PL5 Characterization of
Lanosterol Synthase (Erg7) from Pneumocystis jirovecii. THOMAS M.
SESTERHENN,1 A. GEORGE SMULIAN,1,2 MELANIE T. CUSHION1,2 ; 1
University of Cincinnati College of Medicine, Cincinnati, OH, USA,
2 Cincinnnati Veterans Affairs Medical Center, Cincinnati, OH, USA.
Although the introduction of HAART has resulted in a marked
reduction of Pneumocystis pneumonia (PCP) in AIDS patients,
mortality remains at the same rate today as it was in the pre-HAART
era (10- 15%). The rate is even higher in non-AIDS patients. There
is a critical need for new therapeutic options for PCP due to
problems of toxicity with current drugs and the potential of
emerging resistance to standard therapies. One proposed target for
an alternate Pneumocystis therapy is the sterol biosynthesis
pathway. This pathway produces sterol components essential for
viability in these organisms. One enzyme in the pathway, Erg7, is
responsible for the production of lanosterol, the first sterol
intermediate of the mammalian and fungal sterol biosynthesis
pathways. Recent studies in our laboratory using a combined in
silico discovery platform and in vitro screening system have
identified potential inhibitors to the P. carinii Erg7p (PcErg7).
Although drug responses in animal models have been reflective of
activity in humans, it would be desirable to screen these putative
scaffolds against the P. jirovecii Erg7 (PjErg7). To that end,
portions of a putative PjErg7 gene were amplified by PCR from
clinical isolates using degenerate primers based on the sequence of
Erg7 from P. carinii and several other closely related fungi. The
putative PjErg7 sequence was found to have a high homology to
PcErg7. Additionally, in silico analysis showed a number of
consistencies between the two genes, including conserved catalytic
domains and predicted trans-membrane regions. Previous studies by
our lab and others have established a set of tools that provided
evidence for the function and localization of the PcErg7. We
propose to apply these tools to the putative PjErg7 to see if there
is correlation between the function and localization of PjErg7 and
PcErg7. [Supported by the National Institutes 22. 22 of Health
NIAID N01AI-25467, R01 AI050450 and R01AI44651; and the Department
of Veterans Affairs] PL6 Bone Marrow Failure and Immune Responses
to Pulmonary Pneumocystis (PC) Infection. DAVID TAYLOR, MICHELLE
WILKISON, NICOLE MEISSNER, Department of Veterinary Molecular
Biology, Montana State University, Bozeman, MT 59718, USA. We
recently demonstrated in a mouse model that lack of type I
IFN-signaling in lymphocyte competent mice (IFNAR-/- mice) results
in bone marrow (BM) depression, lymphocyte-deficiency ( RAG-/-
mice) does not significantly affect hematopoiesis. However, lack of
both lymphocytes and IFNAR (IFrag-/- mice) results in BM failure
due to apoptosis of all lineages following Pneumocystis (PC) lung
infection. Infection with other fungal pathogens such as
Cryptococcus did not induce BM failure. BM failure following PC
infection was also associated with loss of bone mass. Here we
further examined mechanisms involved in the induction of BM failure
in our system. We found that lack of IFNAR-signaling during PC lung
infection resulted in increased oxidative stress in BM cells and
increased osteoclast activity, as measured by TRAP assay. Increased
total Caspase activity in BM cells correlated with cell loss.
Extensive kinetic studies revealed that signals at day 7 post
infection appeared critical in determining the outcome of disease.
Quantitative RT-PCR at day 7 demonstrated excessive up-regulation
of iNOS in BM cells from IFrag-/- mice compared to RAG-/- mice,
significantly reduced expression of OPG, a decoy receptor for the
apoptosis-inducing cytokine TRAIL and the
osteoclast-differentiation factor RANKL, and down-regulation of the
anti- apoptotic factor Bcl10, Bcl2, Birc2 (IAP) and others. Caspase
8 activity, as the initiator caspase for the extrinsic pathway of
apoptosis, was induced in BM cells of both IFrag-/- and RAG-/- mice
following PC lung infection. However, only in IFrag-/- mice was the
executioner caspase 3 also induced which is followed by apoptosis.
Conclusion: We propose that lack of type-I-IFN-signaling negatively
affects the regulation of oxidative stress, decoy mechanisms for
TRAIL and RANKL in BM cells as well as anti-apoptotic mechanisms.
This may have accelerated BM cell death following systemic
responses to PC lung infection resulting in BM failure due to
excessive induction of Capase activity via the extrinsic pathway of
apoptosis. Support: RO1HL090488 and COBRE 2P20RR020185- 06 PO1 Cell
Wall Assembly Components of Pneumocystis carinii Exhibit Unique
Activity in Response to Alterations in Environmental Temperature.
DEANNE M. HEBRINK1 , THEODORE J. KOTTOM1 , ANDREW H. LIMPER1 ; 1
Thoracic Diseases Research Unit, Mayo Clinic, Rochester, MN, USA.
Pneumocystis carinii is an opportunistic organism that causes
pneumonia in immune- compromised hosts. Recent studies in our lab
have reported interesting responses in P. carinii life cycle
proteins to alterations in environmental temperature (Burgess et
al., Am J Resp Cell Molec Biol 2009). The purpose of this study was
to investigate the effects of variations in environmental
temperature the organism may encounter on components of the
cell-wall assembly machinery such as 1,3- -glucan synthase, which
is involved in synthesizing the main structural component of the
cell wall, namely 1,3--glucan. The activity of the 1,3--glucan
synthase enzyme was determined at incubation temperatures ranging
from 4o C to 45o C by measuring 23. 23 incorporation of [14
C]-uridine 5- diphosphoglucose (UDP-glucose), the substrate for the
enzyme into insoluble carbohydrate. Interestingly, these
experiments revealed the enzyme is most active at temperatures
between 4o C and 25o C and dramatically lower at physiologically
relevant temperatures such as 37o C. Real-time PCR data further
confirmed a similar temperature response at the transcript level
for PcGsc-1 with greater RNA expression at 4o C and 10o C compared
to physiological temperatures. RNA expression for transcripts that
encode other proteins involved in cell-wall biogenesis such as
PcCdc42, PcMsg1, and PcKre6 also have higher RNA expression at sub-
physiological temperatures while RNA expression for other
regulatory genes including PcCdc13 and PCFlo8 do not seem to be
influenced by changes in environmental temperature. Lastly, future
aims will determine whether environmental temperature has any
effect on the overall metabolic activity of P. carinii by measuring
cellular ATP production in Pneumocystis after short-term culture at
these temperatures. These studies suggest that temperatures more
typically found outside of the mammalian host are more conducive to
processes involved in cell-wall assembly and provide interesting
insights into the life cycle of P. carinii. PO2 Influence of
Climate and Ambient Air Pollutants on Pneumocystis jirovecii
Pneumonia (PcP) Hospital Admission and on Antibody Responses to
Major Surface Glycoprotein (Msg) in HIV-Infected Patients from San
Francisco. K. DJAWE, MS1 , L. HUANG, MD2 , K.R. DALY, PHD1 , L.
LEVIN, PHD1 , J. KOCH1 , A. SCHWARTZMAN, BS2 , S. FONG, BA2 , B.
ROTH, MPH2 , A. SUBRAMANIAN, MD2 , K. GRIECO, DO2 , L. JARLSBERG,
BA2 , P.D. WALZER, MD, MSC1 . 1 VAMC and U. Cincinnati, Cincinnati,
OH, United States, 2 San Francisco General Hospital (SFGH) and U.
California SF, San Francisco, CA, United States. Background:
Pneumocystis jirovecii pneumonia (PcP) is an important
opportunistic infection in immunocompromised patients. The
influence of seasonality on this fungal infection is not well
established. Furthermore, the influence of ambient air pollutants
on PcP incidence is unknown. The Major Surface Glycoprotein (Msg)
is a crucial protein complex in Pneumocystis pathogenicity and is
involved in host-organism interaction. Different risk factors have
been associated with antibody responses to recombinant Msg
fragments, but the influence of climate and air pollutants on
antibody responses to Msg fragments is unknown. Objectives: The
objectives of this study are to determine the influence of climate
and ambient air pollutants on PcP incidence and on antibody
responses to Pneumocystis Msg recombinant fragments. Methods: From
January 1, 2000 to December 31, 2008, 140 HIV+ patients were
admitted at San Francisco General Hospital (SFGH) with confirmed
PcP. Daily climate data from San Francisco were collected from the
California Irrigation Management Information System (CIMIS)
website, and daily pollutants data were collected from San
Francisco EPA website. Logistic regression was used to determine
the risk of PcP associated with different environmental parameters
at the time of hospital admission and at different lag times (29-31
days and 59-61 days before admission). Tobit regression was used to
estimate the effect of climate and air pollutants on antibody
responses to the Msg fragments. Results: We found that PcP hospital
admission was associated with higher temperature and higher ozone
levels, but only the effect of temperature was significant (P =
0.02). In contrast, lower humidity, rainfall, CO, NO2 and SO2 was
insignificantly associated with an increase in PcP admission. 24.
24 Only lower lag humidity 29-31 days was significantly associated
with an increase in PcP admission (P = 0.01). None of the
parameters had a lag 59-61 days significant effect on PcP. We also
found significant seasonal variations in antibody responses to
MsgA, the amino terminus fragment (P = 0.01), but not to MsgC, the
carboxyl terminus fragment. Both temperature and CO levels had
significant effects on antibody responses to MsgA, but only
humidity had a significant effect on antibody responses to MsgC
when controlling for CD4 cell counts. Conclusion: Thus, climate and
ambient air pollutants have complex effects on the occurrence of,
and antibody responses to PcP, in San Francisco. Further
investigation of these factors should help in understanding of the
epidemiologic features of PcP. PO3 Pneumocystis jirovecii
Colonization in Patients Undergoing Treatment with the TNF-
Antagonist Agent Infliximab. RUBN MORILLA1 , ISABEL MARTN- GARRIDO1
, GUSTAVO WISSMANN2 , VICENTE FRIAZA1 , NIEVES RESPALDIZA1 , RAFAEL
TERAN1 , MARIA T. MARTINEZ-RISQUEZ1 , ELENA CAMPANO1 , FRANCISCO J.
MEDRANO1 , CARMEN DE LA HORRA1 , JOS M. VARELA1 , JUAN POVEDANO3 ,
ENRIQUE J. CALDERN1 ; 1 Instituto de Biomedicina de Sevilla and
CIBER de Epidemiologia y Salud Pblica. Service of Internal Medicine
Virgen del Rocio University Hospital, Seville, Spain; 2 Grupo de
Estudos em Pneumocystis, Servio de Infectologia, Hospital de
Clnicas de Porto Alegre, Brasil; 3 Service of Rheumatology, Virgen
del Roco University Hospital, Seville, Spain. Infliximab, a
chimeric anti-tumor necrosis factor (TNF)- monoclonal antibody, has
become an established effective therapy for inflammatory rheumatic
disease. However, TNF- is a critical factor in host defense and the
suppression of its biological activity may be associated with the
increased risk of opportunistic infections. The frequent use of
infliximab in clinical practice has identified Pneumocystis
jirovecii pneumonia (PcP) as a serious complication. Individuals
colonized with Pneumocystis may be at high risk of development of
PcP when they have undergone immunosuppressive therapies. Hence, we
addressed the question of the frequency of Pneumocystis
colonization among patients treated with infliximab. We examined
125 oropharyngeal washes (OW) collected from 78 individuals with
rheumatoid arthritis, 30 with ankylosing spondylitis and 17 with
psoriatic arthritis. Half of them had undergone infliximab therapy.
Using a real- time polymerase chain reaction assay based on the
amplification of the large subunit of mitochondrial DNA (mtLSU
rDNA) of Pneumocystis, P. jirovecii colonization was detected in 32
(25,6%) patients. Colonization was associated with use of
corticosteroid and methotrexate, but only duration of infliximab
therapy was significantly and independently associated with
Pneumocystis colonization in a multivariate regression model. There
is a high rate of P. jirovecii colonization among patients with
rheumatologic diseases treated with infliximab. The identification
of patients colonized by P. jirovecii before starting the treatment
with infliximab using noninvasive samples as OWs could be a
strategy for PcP prevention that warrants further investigation for
its validation. PO4 Dihydropteroate Synthase Mutations in
Pneumocystis jirovecii Isolated from Patients with Pneumocystis
Pneumonia in Brazil. GUSTAVO WISSMANN1 , ROSECLER B. MENDES1 ,
VICENTE FRIAZA2 , ANDR L. MLLER1 , RUBEN 25. 25 MORILLA2 , CARMEN
DE LA HORRA2 , LUCIANO Z. GOLDANI1 , ENRIQUE J. CALDERN2 ; 1 Grupo
de Estudos em Pneumocystis, Servio de Infectologia, Hospital de
Clnicas de Porto Alegre, Brasil; 2 Instituto de Biomedicina de
Sevilla and CIBER de Epidemiologia y Salud Pblica. Virgen del Rocio
University Hospital, Seville, Spain. Pneumocystis pneumonia (PcP)
is a common opportunistic infection causing morbidity and mortality
in immunocompromised patients, particularly in HIV-infected
individuals. A combination of trimethoprim (TMP) and
sulfamethoxazole (SMX) is the first choice for both prophylaxis and
treatment of PcP. Several studies have showed an association
between the failure of sulfa prophylaxis and the presence of
mutations in P. jirovecii dihydropteroate synthase (DHPS) gene,
especially at nucleotide positions 165 and 171 which entail an
amino acid change at positions 55 and 57. Bronchoalveolar lavage
samples were obtained from 11 PcP patients from January to
September 2007 at the Hospital de Clnicas de Porto Alegre, Brazil.
The DNA was extracted using a commercial kit (Qiagen, Hilden,
Germany). A touchdown PCR using the primers DHPS-3 (5-GCG CCT ACA
CAT ATT ATG GCC ATT TTA AAT C-3) and DHPS-4 (5-GGA ACT TTC AAC TTG
GCA ACC AC-3) was used to amplify the DHPS gene. DHPS
polymorphisms, at codons 55 and 57, were detected by using AccI and
HaeIII restriction enzymes. The P. jirovecii DHPS gene was
amplified successfully in samples from 9 of 11 patients (82%). DHPS
wild type 55/Thr, 57/Pro were detected in 7 patients, and mutations
were obtained in two patients (one had a single mutation 55/Thr,
57/Ser and the other had a double mutation 55/Ala, 57/Ser). The
absence of P. jirovecii DHPS mutations was reported from Brazilian
PcP patients during 1997-2004. The present study could detect
mutant genotypes in samples collected during 2007 in Brazil. As PcP
is a common opportunistic infection in patients with AIDS in
developing countries, it will be important to monitor the
prevalence of P. jirovecii DNA mutations related to resistance to
sulfa-drugs in these countries. PO5 DHPS Mutations in Pneumocystis
jirovecii Isolated from Patients with PcP in Santiago, Chile.
CAROLINA A. PONCE,1 MAGALI CHABE,2 CLAUDIO GEORGE,1 ALEJANDRA
CARDENAS,1 LUISA DURAN;2 JULIA GUERRERO,1 LAURENCE HUANG,3 ROBERT
F. MILLER,4 SERGIO L. VARGAS1 ; 1 University of Chile School of
Medicine, Santiago, Chile, 2 Institute Pasteur du Lille, France, 3
University of California San Francisco, San Francisco, CA, USA, 4
University College London, UK. Trimethoprim-sulfamethoxazole (TMP-
SMZ) is the mainstay of therapy and prophylaxis for P. jirovecii
pneumonia (PcP). Concern about emerging resistance has been raised
because mutations in the fas gene coding for the dihydropteroate
synthase (DHPS) enzyme, a target for sulfa drugs, has been
described in 4-81% of P. jirovecii isolates in different parts of
the world. P. jirovecii isolates from 94 patients (median age, 39
years; range, 5-82 years) whose respiratory samples were referred
to our laboratory in Santiago, Chile for diagnostic analysis
between 2002 2010, and confirmed as PcP by Gomori-Grocott stain and
fluorescent-labeled monoclonal antibody stain or PCR, were
evaluated for mutations at positions 165 and 171 using Touch Down
PCR with primers DHPS-3 and DHPS-4 and restriction enzymes Accl and
Hae lll for RFLP-PCR analyses. Mutations were classified according
to the pattern of band polymorphisms visualized by UV in 2% agarose
gel with ethidium bromide. Fas gene 26. 26 polymorphisms were
detected in 43 (45.7%) of the 94 patients. Five (5.3%) were found
to have a single mutation at position 165; one (1.1%) had a single
mutation at position 171. Fifteen (16.0%) had double mutations at
positions 165 and 171. Twelve (12.8%) patients were found to have
co-infections with wild type and a mutation at position 165; and 10
(10.6%) had wild type plus double mutations. DHPS mutations are
frequent in Chile and their relationship with clinical outcome of
PcP needs to be explored especially since TMP-SMZ is the only
therapeutic option in our country. [Fondecyt 1060750] PO6
Characterization of Mouse Alveolar Macrophage Binding to
Pneumocystis murina Cyst and Trophic Forms. A.D. ASHBAUGH, M.J.
LINKE, M.T. CUSHION; Veterans Affairs Medical Center, Cincinnati,
OH, and University of Cincinnati College of Medicine, Cincinnati,
OH. Alveolar macrophages are the primary phagocytic cells in the
lung and are the principal effector cells involved in clearance of
Pneumocystis infection. Several alveolar macrophage receptors have
been shown to be involved in Pneumocystis binding and uptake
facilitated by Pneumocystis glycoproteins and -1,3-D-glucan.
However, it is unknown whether all life cycle forms of Pneumocystis
are bound and taken up by alveolar macrophages and if so, whether
different receptors are used for each form. Echinocandin treatment
(ECH) of Pneumocystis-infected mice has been shown to shift the
population to one made up of almost exclusively trophic forms. The
ECH mouse model was used to interrogate the binding and
phagocytosis of P. murina by alveolar macrophages. Alveolar
macrophages from mouse lungs were adhered to 10 well glass slides
at a concentration of 104 macrophages/well. Fluorescently labeled
mixed populations of P. murina from control mice or enriched
trophic populations from ECH mice were added to the macrophages,
incubated and attachment assessed by fluorescent microscopy.
Results were expressed as the percentage of alveolar macrophages
with at least one 1 attached organism. Preliminary data showed
binding averages of 55% (36% cysts and 19% trophs) and 49% (33%
cysts and 16% trophs) when P. murina concentrations of 4 x 105 and
2 x 105 organisms/well were used, respectively. When the ECH P.
murina were used at the same concentrations, the binding averages
were reduced to 36% (5% cysts and 31% trophs) and 31% (5% cysts and
26% trophs) respectively. This data suggests a higher affinity of
cyst binding to alveolar macrophages than trophs due to their more
complex surface antigen profile. Further experiments are needed to
better understand the fate of each life cycle form in alveolar
macrophage mediated clearance. PO7 Human Pathogenic Microsporidia:
Detection and Genotyping in HIV-positive and negative patients from
Portugal. M. L. LOBO 1 , L. XIAO2 , F. ANTUNES 3 , O. MATOS1 ; 1
Instituto de Higiene e Medicina Tropical, CMDT, UNL, Lisboa,
Portugal, 2 Centers for Disease Control and Prevention, Atlanta,
GA, USA, 3 Hospital de Santa Maria, FM/UL, Lisboa, Portugal. Most
cases of microsporidiosis are associated with severe depletion of
the patients immune system. The main goal of this study was to
provide information on the occurrence of microsporidia involved in
humans in Portugal, and evaluate their public health importance.
Stool from 856 children and adults (561 HIV-positive, 255 HIV-
negative, 29 persons with undetermined HIV 27. 27 status, and 9
with other immunosuppressions) with gastrointestinal complains,
urine from 50 patients (40 HIV-positive and 10 HIV- negative), and
pulmonary specimens from 200 patients (150 HIV-positive and 50 HIV-
negative) were analysed for the presence of microsporidia by PCR.
The presence of Enterocytozoon bieneusi and the Vittaforma- like
parasite were detected in 6.3% (54/856) and 6.8% (58/856) of stool
samples, respectively. A statistically significant association was
observed between microsporidia infection and age; children were at
a significantly higher risk of infection than adults.
Encephalitozoon intestinalis was identified in 2% (1/50) of urine
samples. Encephalitozoon cuniculi and Vittaforma-like parasite were
each identified in 0.5% (1/200) of the pulmonary specimens studied.
Genotyping analysis confirmed the presence of pathogenic
microsporidia in Portuguese patients. Data from the study showcase
the harmful role of microsporidia in susceptible populations.
(Supported in part by Associao para a Investigao e Desenvolvimento
da Faculdade de Medicina de Lisboa. ML Lobo is supported by PhD
grant-SFRH/BD/34674/2007-FCT) PO8 QSAR Study of New Inhibitors for
Pneumocystis carinii Oxidosqualene Cyclase. ALEKSEY POROLLO1 ,
MARGARET S. COLLINS1,2 , MELANIE T. CUSHION1,2 ; 1 University of
Cincinnati College of Medicine Department of Environmental Health,
Cincinnati, OH, USA; 2 University of Cincinnati Department of
Internal Medicine and the Cincinnati Veterans Affairs Medical
Center, Cincinnati, OH Pneumocystis pneumonia (PcP) remains a
significant opportunistic infection in patients with compromised
immune systems. The limited repertoire of treatment alternatives
and emerging resistance to the standard anti-PcP therapies drives
the search for new chemotherapeutic agents. In this work, we
focused on the oxidosqualene cyclase (OSC) enzyme in the sterol
biosynthesis pathway using Pneumocystis carinii (Pc) as a model
organism. In contrast to the efforts to identify specific
inhibitors for its active site, we evaluated another structural
cavity as an alternative target. This structural site is involved
in OSC enzymatic function and contains amino acids conserved among
the fungi. Using large scale virtual screening, three new classes
of inhibitors for OSC were identified. An array of compounds,
derivatives of these classes, was experimentally validated using an
in vitro drug screening system for Pc. A cytotoxicity assay to
determine potential toxicity was conducted using the mammalian cell
lines, A549 (human) and L2 (rat). Here, we report results of the in
vitro assays and corresponding observed quantitative structure-
activity relationships (QSAR). Of 35 total compounds evaluated, 3
compounds were found to show marked anti-Pc activity, 4 and 12 with
moderate and slight activity, respectively. The active compounds
exhibit various levels of cytotoxicity, including 2 inhibitors of
the desired efficacy with 1:10 activity/toxicity ratio. [Supported
by the Midwest Center for Emerging Pathogens, Cincinnati OH;
National Institute of Environmental Sciences NIEHS P30- ES006096;
National Institutes of Health NIAID N01AI-25467, R01 AI050450 and
R01AI44651; and the Department of Veterans Affairs]. PO9 Factors
Influencing the Carriage, Colonization, and Transmission of
Pneumocystis carinii. K.A. LYNCH, and M.T. CUSHION, University of
Cincinnati College of Medicine and the Cincinnati 28. 28 Veterans
Affairs Medical Center, Cincinnati, Ohio, USA. Studies suggest that
Pneumocystis can exist with little consequence to hosts with intact
immune systems, although the length of resident time within the
lung is not known nor the host factors that may influence
colonization in hosts with intact immune systems. Likewise, the
widespread prevalence of Pneumocystis in these immune competent
(IC) populations suggests an efficient mechanism of transmission.
In the present study we evaluated the effects of gender and age of
IC rats on length of P. carinii (Pc) colonization and ability to
transmit the infection. In the 1st study, the period of carriage of
Pc in IC male and female rats at different ages (weanling,
juvenile, adult) was determined after intra-tracheal inoculation of
2 x 107 Pc followed by PCR with Rc1 and Rc2 primers of oral swabs
taken for up to 1 year. Once the rats had 3 consecutive negative
oral swabs they were sacrificed and their lung tissue was processed
and verified for lack of Pc by PCR. Time to clearance ranged from
169 d to 190 d with no significant differences among the groups,
although weanling and juvenile females trended towards earlier
clearance than males. In the 2nd study, adult IC male and female
rats were inoculated as above, and immunosuppressed at 2-, 4- and 6
months post inoculation to determine whether Pc were infective to
the host after short to long periods of carriage. Rats
immunosuppressed after 2 months of carriage produced significantly
more infections in 5/8 (female) and 6/8 (male) rats than rats after
4-6 months of carriage (0-2 rats per group), without difference
between genders. In the 3rd study, the influence of age on the
transmission in IC rats was evaluated. IC intra-tracheally
inoculated donor weanling, juvenile and adult female rats were
housed with age-matched uninoculated IC female recipients for a
total of 14 pairs. Oral swabs were taken on a daily basis up until
14 days of exposure. Seven matched pairs in each group that were
positive by 7-days exposure were removed, separated, and
immunosuppressed. The remaining 7 pairs were co-housed for 14 days,
separated and immunosuppressed. Juvenile rats were the first to
become positive for Pc with all 7 of the early group turning
positive in 2 days. Weanlings took an average of 7 days of exposure
and adults 9 days of exposure. However, resultant infections were
generally higher in the adult recipients. Interestingly, with 1
exception, the recipient rats had higher burdens than the donor
rats. Under the conditions described here, gender does not appear
to play a role in clearance of Pc in IC rats. IC rats infected with
Pc were efficient in transmitting the infection to IC recipients,
but the time in which Pc in IC rats that could produce an active
infection was limited to less than 4 months, suggesting there is a
constant circulation of Pc in IC populations. [Supported by
National Institutes of Health NIAID N01AI-25467, R01 AI050450 and
R01AI44651 and by the Medical Research Service, Department of
Veterans Affairs] PO10 Microscopic Studies of Anncaliia algerae-
Infected Cell Cultures and Analysis of its Beta-Tubulin Gene
Suggest Albendazole Sensitivity. MARIANITA SANTIANA, PETER
TAKVORIAN, CYRILLA PAU, ANN CALI. Rutgers University, Newark, NJ,
USA. The microsporidium, Anncaliia algerae (Brachiola algerae), is
an obligate intracellular parasite. It has been identified as an
opportunistic human pathogen but treatment has not been evaluated
for infection with this organism. Albendazole has been the
medication of choice used to treat other microsporidial infections
affecting humans, with varying results. We have studied various
parameters of albendazole treatment during infection in rabbit
kidney cells and have found 29. 29 that treatment causes
abnormalities to the structure of the intracellular stages of the
parasite. Sustained albendazole treatment also has an attenuating
effect on infection, inhibiting up to 98% of spore production, but
release from treatment re-establishes the infection without new
exposure to the parasite. In addition, we also performed an
analysis of the beta-tubulin gene sequence and found that 5 of the
6 specific amino acids, which are highly suggestive of
benzimidazole sensitivity, are conserved among Anncaliia algerae
and other organisms sensitive to albendazole. Results from this
study give an insight to a possible long-term treatment for
Anncaliia algerae infections that can be affordable and widely
available. PO11 Cryptosporidium spp. in Pet Birds in Henan, China:
Prevalence and Molecular Characterizations. MENG QI,1 RONGJUN
WANG,1 CHANGSHEN NING,1 LONGXIAN ZHANG,1 FUCHUN JIAN,1 YANRU SUN,1
LIHUA XIAO2 ; 1 The College of Animal Science and Veterinary
Medicine, Henan Agricultural University, Zhengzhou 450002, China, 2
Division of Foodborne, Bacterial, and Mycotic Diseases, National
Center for Emerging and Zoonotic Infectious Diseases, Centers for
Disease Control and Prevention, Atlanta, GA 30333, USA To
characterize the prevalence of Cryptosporidium species/genotypes in
pet birds in Henan, China, 434 fecal samples were acquired from 14
families of birds in pet shops. The overall prevalence of
Cryptopsoridium was 8.1% (35/434) by the Sheathers sugar flotation
technique. The Cryptosporidium-positive samples were analyzed by
DNA sequence analysis of the small subunit (SSU) rRNA gene. Three
Cryptosporidium species and two genotypes were identified,
including C. baileyi (18/35 or 51.4%) in five red-billed
leiothrixes (Leiothrix lutea), four white Java sparrows (Padda
oryzivora), four common mynas (Acridotheres tristis), two zebra
finches (Taeniopygiagutttata), a crested Lark (Galerida cristata),
a Gouldian finch (Chloebia gouldiae), and a black-billed magpie
(Pica pica); Cryptosporidium meleagridis (3/35 or 8.6%) in a
Bohemian waxwing (Bombycilla garrulus), a Rufous turtle dove
(Streptopelia orientalis), and a fan- tailed pigeon (Columba
livia); Cryptosporidium galli (5/35 or 14.3%) in four Bohemian
waxwings (Bombycilla garrulus) and a silver-eared Mesia (Leiothrix
argentauris); Cryptosporidium avian genotype III (3/35 or 8.6%) in
two cockatiels (Nymphicus hollandicus) and a red-billed blue magpie
(Urocissa erythrorhyncha); and Cryptosporidium avian genotype V
(6/35 or 17.1%) in six cockatiels (Nymphicus hollandicus). Among
the pet birds, 12 species represented new hosts for Cryptosporidum
infecitons. This is the first report of cryptosporidiosis in pet
birds in China. PO12 Standardization of PCR Primers for gp60-based
Subtyping of Cryptosporidium hominis and Cryptosporidium parvum.
SATOMI KATO, LIHUA XIAO, Atlanta Research and Education Foundation,
Centers for Disease Control and Prevention. The 60 kDa glycoprotein
(gp60 or cpgp15/45) gene has been widely used for subtyping
human-pathogenic Cryptosporidium spp. However, there are no
standardized primers for PCR analysis and poor PCR sensitivity has
been a major issue associated with the gp60 subtyping. In this
study, we assessed the specificity and sensitivity of 43 published
primers. Primers with narrow specificity were eliminated based on
sequence polymorphism in the primer region identified 30. 30 with
an alignment of nucleotide sequences from major subtype families of
C. hominis and C. parvum. Based on this, 10 forward and 9 reverse
primers were further evaluated using DNA preparations of major
subtype families of C. hominis (Ia, Ib, Id, and Ie) and C. parvum
(IIa, IIc, and IId) and real-time PCR. Three outer primer
combinations, gp15ATG/HW4, HW3/HW4, and S60.ATGF/gp15extR, and two
inner primer combinations, AL3532/LX0375 and AL3532/AL3535,
produced better amplifications. These primers were able to amplify
DNA of all major subtype families of C. hominis and C. parvum.
Based on amplification efficiency in real-time PCR, the combination
of S60.ATGF/gp15extR in primary PCR and AL3532/AL3535 in secondary
PCR was selected. The use of standardized gp60 primers should
improve the gp60-based subtyping and promote its use in molecular
epidemiologic investigations of cryptosporidiosis. PO13 Preliminary
Biochemical Data on a Type II Thioesterase from Cryptosporidium
parvum (CpTEII). FENGGUANG GUO, GUAN ZHU; Department of Veterinary
Pathobiology, Texas A&M University, TX, USA. The genome of
Cryptosporidium parvum encodes a type II thioesterase (TEII)
ortholog. This is unique to Cryptosporidium as TEII is absent in
other apicomplexans. To understand its potential role in the
parasite, we cloned the CpTEII gene and expressed its product as
maltose-binding protein (MBP)-fused protein for biochemical
analysis. We have tested its activity towards acyl-CoAs with
various chain lengths (C4 to C24) and discovered that CpTEII is
only able to hydrolyze C6- to C12-length acyl-CoAs. It displays the
highest activity towards decanoyl CoA. The preliminary biochemical
data is in line with our hypothesis that CpTEII might play a
similar role as in bacteria, i.e., the removal of aberrant acyl
chains from polyketide synthases (PKSs). We are currently studying
its activity towards acyl-acyl carrier protein (ACP) derived from
C. parvum fatty acid synthase (CpFAS1) and polyketide
synthase(CpPKS1) to further delineate the potential involvement of
CpTEII in the fatty acid/polyketide synthesis in the parasite. PO14
Hospital-Based Monitoring of Cryptosporidium parvum, Giardia
lamblia and Entamoeba histolytica in the Republic of Korea,
2004-2008. HYENG-IL CHEUN*, YI-YOUNG LIM, SHIN- HYEONG CHO,
JUNG-WON JU, SANG- EUN LEE, JEONG-YEON KIM, WON-JA LEE. Division of
Malaria and Parasitic Diseases, National Institute of Health, Korea
Centers for Disease Control and Prevention, Seoul 122-701, Korea.
Infectious status of Cryptosporidium parvum, Giardia lamblia and
Entamoeba histolytica as water- and food-borne parasite was
investigated in hospital-based diarrheal patients in the Republic
of Korea from 2004 to 2008. A total of 104,483 stool samples were
collected from the across nation in 106 hospitals. Infections in
different age groups, gender and seasonal distribution was
analyzed. Fecal samples that were positive for C. parvum, G.
lamblia and E. histolytica were 0.5, 0.5 and 0.3%, respectively.
There was not significant difference by gender for C. parvum (P =
0.3748), G. lamblia (P = 0.1796) and E. histolytica (P = 0.8362).
If those over 60- years old was taken as the reference group,
positivity of 2-9 years olds (P = 0.0019) was significantly higher
in C. parvum, however all age group were lower than the over-60 age
group with respect to infection with G. lamblia and E. histolytica.
Analyses of infections during certain years showed that C. 31. 31
parvum and E. histolytica infections were highest during the
2004-2005 period, however G. lamblia infections were significantly
lower during 2004-2005 compared to other years. Analyses of
seasonality, C. parvum and G. lamblia infections peaked during
March and September, and E. histolytica infections peaked in June.
Information on hospital-based protozoa in terms of patient gender,
age, and seasonal patterns would improve diarrheal medical care,
reduce the burden of acute gastrointestinal infections and help the
development of control strategies for diarrheal diseases in the
Republic of Korea. PO15 Low Prevalence of Pneumocystis Pneumonia
(PCP) but High Prevalence of Pneumocystis Dihydropteroate Synthase
(DHPS) Gene Mutations in HIV-Infected Persons in Uganda. LAURENCE
HUANG, MD1 , STEVE M. TAYLOR, MD, MPH2 , WILLIAM WORODRIA, MBCHB,
MMED3 , LEAH JARLSBERG, BA1 , J. LUCIAN DAVIS, MD1 , ADITHYA
CATTAMANCHI, MD1 , SAMUEL D. YOO, MD3 , ALFRED ANDAMA, BS3 , SASKIA
DEN BOON, PHD3 , RACHEL KYEYUNE, MBCHB3 , STEVEN MESHNICK, MD, PHD2
. 1 San Francisco, CA, USA, 2 Chapel Hill, NC, USA, 3 Kampala,
Uganda. Rationale: PcP was the most frequent cause of pneumonia in
hospitalized HIV-infected persons in Uganda who had negative sputum
acid fast bacillus (AFB) smears and underwent bronchoscopy,
accounting for 39% (32 of 83) of cases [Worodria 2003]. Although
trimethoprim-sulfamethoxazole (TMP-SMX) is effective in preventing
PcP, its use has been associated with the presence of DHPS
mutations and putative TMP-SMX drug resistance. As TMP-SMX use has
increased in Uganda, the current prevalence of PcP and DHPS
mutations are unclear. Objectives: 1. To determine the prevalence
of PcP in hospitalized HIV-infected persons with suspected
pneumonia and negative sputum AFB smears. 2. To determine the
prevalence of DHPS mutations in persons with PcP. Methods: We
enrolled consecutive HIV- infected adults with cough >2 weeks
and suspected pneumonia who were admitted to Mulago Hospital in
Kampala, Uganda between September 2007 and July 2008. Patients
underwent standardized evaluation including sputum x2 sent for AFB
smear and bronchoscopy with bronchoalveolar lavage (BAL) if AFB
smears were negative. BAL specimens were examined for PcP at Mulago
using a modified Giemsa stain and also sent to the University of
North Carolina for polymerase chain reaction (PCR) and DNA
sequencing at the DHPS locus. Results: Pneumocystis was identified
microscopically in 7 of 133 (5.3%) BAL specimens. Overall, persons
with PcP had a lower median CD4+ T-cell count (10 cells/ul vs. 86
cells/ul, P = 0.01) and tended to be less likely to have received
PcP prophylaxis or antiretroviral therapy than persons without PcP.
There was 100% concordance between modified Giemsa microscopy and
PCR. DHPS PCR was positive in all 6 of the PcP microscopy- positive
BAL specimens tested and negative in all 124 of the PcP
microscopy-negative specimens tested. All 6 PCR-positive BAL
specimens contained Pneumocystis DHPS mutations; the most frequent
DHPS genotype was GT3 (Thr55/Ser57), seen in 4 of the 6 cases.
Conclusions: The prevalence of PcP in hospitalized HIV-infected
adults undergoing bronchoscopy at Mulago Hospital is low and has
decreased compared to an earlier study (October 1999 through
February 2000) conducted prior to the widespread use of TMP-SMX for
PcP prophylaxis in Uganda. The prevalence of Pneumocystis DHPS
mutations is high. Interestingly, the most frequent DHPS genotype
(GT3) reported in this study is rarely reported in the US and has
32. 32 been associated with the use of sulfadoxine (plus
pyrimethamine) rather than sulfamethoxazole (TMP-SMX). This raises
the question of whether sulfa drugs used for indications other than
PcP prophylaxis such as malaria treatment may also be associated
with the presence of PcP that contains DHPS mutations. PO16
Variation in the erg11 Gene from Pneumocystis jirovecii. SCOTT P.
KEELY,1 JAMES R. STRINGER,1 CARLO CONTINI,2 BETTINA LUNGREN,3
RONALD BRUBAKER4 , LAURA Q. JOHNSTON,5 EDNA S. KANESHIRO5 ; 1 Dept.
Molecular Genetics, Biochemistry & Microbiology, 5 Dept.
Biological Sciences, Univ. Cincinnati, Cincinnati, OH; 2 Dept.
Clinical & Experimental Medicine, Univ. Ferrara, Ferrara,
Italy, 3 Dept. Clinical Microbiology, Hvidovre Univ. Hospital,
Hvidovre, Denmark. 4 Department of Pathology, Christ Hospital,
Cincinnati, OH. The erg11 gene codes for sterol 14- demethylase
(14DM), a key enzyme in sterol biosynthesis and the target for
triazole antimycotic drugs. Two sterol composition phenotypes have
been described in patients with pneumonia caused by P. jirovecii;
one phenotype has a much higher amount of lanosterol derivatives
(C31 and C32 sterols) dominated by pneumocysterol. Most of the
Pneumocystis-distinct sterols in this phenotype were those with a
methyl group present at the C-14 position of the sterol nucleus
suggesting that the P. jirovecii present might lack 14DM activity.
To gain insight into whether the apparent lack of 14DM activity
might be due to mutation of the erg11 gene, we have analyzed a
1,000-bp segment of this 1,800-bp gene in nine specimens of P.
jirovecii (from the USA, Italy and Denmark). Thus far, two alleles
of the erg11 gene have been observed: allele 1 with G at position
238 and allele 2 with A at this nucleotide position. All three
specimens from Italy had allele 2, two of which were shown to
contain high proportions of pneumocysterol. The other specimens
analyzed had allele 1. The two alleles differ at a single site that
is located in an apparent intron. Sequencing of the entire erg11
gene from these and additional samples is in progress [Supported in
part by NIH grant RO1 AI064084]. PO17 Development of a Multilocus
Sequence Typing Tool for Cryptosporidium muris and Cryptosporidium
andersoni. YAOYU FENG,1 WENLI YANG,2 UNA RYAN,3 LONGXIAN ZHANG,4
MARTIN KV,5 BETISLAV KOUDELA,5 NA LI,2,6 RONALD FAYER,7 LIHUA XIAO2
; 1 East China University of Science and Technology, Shanghai,
China, 2 Centers for Disease Control and Prevention, Atlanta, GA,
USA, 3 Murdoch, Western Australia, Australia, 4 Henan Agricultural
University, Zhengzhou, China, 5 Academy of Sciences of the Czech
Republic, esk Budjovice, Czech Republic, 6 Tongji University,
Shanghai, China,7 U.S. Department of Agriculture, Beltsville,
Maryland, USA. Although genotyping tools have been developed and
widely used in the characterization of the transmission of
intestinal Cryptosporidium spp., they are not available for C.
muris and C. andersoni, two most common gastric Cryptosporidium
spp. of mammals. In this study, we screened the C. muris whole
genome sequencing data for microsatellite and minisatellite
sequences. Among the 13 potential loci (six microsatellite and
seven minisatellite loci) evaluated by PCR and DNA sequencing, four
were eventually chosen. DNA sequence analyses of 27 C. muris and 17
C. andersoni DNA preparations showed the presence of 5-10 subtypes
of C. 33. 33 muris and 1-4 subtypes of C. andersoni at each locus.
Altogether, 11 C. muris and seven C. andersoni multilocus sequence
typing (MLST) subtypes were detected among the 16 C. muris and 12
C. andersoni specimens successfully sequenced at all four genetic
loci. In all analyses, the C. muris isolate (TS03) originated from
an East African mole rat (Tachyoryctes splendens) differed
significantly from other C. muris isolates, approaching the extent
of genetic differences between C. muris and C. andersoni. Thus, a
MLST technique was developed for high resolution typing of C. muris
and C. andersoni. It should be useful in the characterization of
the population genetics and transmission of gastric Cryptosporidium
spp. PO18 Amoebicidal and Amebastatic Activity In Vitro of the
Resin of Gymnosperma glutinosum Against Acanthamoeba castellanii
trophozoites. RODRIGUEZ- MONROY MA1 , PEA-JUAREZ MC2 , OMAA-MOLINA
M1 , GONZALEZ- ROBLES A3 , SALAZAR-VILLATORO L3 , CANALES-MARTINEZ
MM4 .1 Profesor Carrera de Medicina, U.N.A.M. F.E.S. Iztacala. 2
Profesor Carrera de Biologa, U.N.A.M. F.E.S. Iztacala. 3
Departamento de Infectmica y Patognesis Molecular, CINVESTAV,
I.P.N., 4 Profesor Carrera de Medicina, U.N.A.M. F.E.S. Iztacala,
Edo. Mexico. We investigated the in vitro activity of the resin of
Gymnosperma glutinosum (a plant that belongs to the Asteraceae
family, considered one of the most important families of plants in
traditional medicine in Mexico) on Acanthamoeba castellanii
trophozoites. We measured the effects on cell viability by the
reduction of tetrazolium salts (MTT protocol), in an attempt to
identify other useful agents that can be used against Acanthamoeba
sp. Strains of Acanthamoeba sp. constitute a factor contributing to
the occurrence of chronic granulomatous amoebic encephalitis,
keratitis, pneumonia, as well as inflammations of other organs.
Treatment of these diseases is very difficult and not always
effective. A majority of these infections have been fatal. The aim
of this study was to investigate and evaluate the in vitro effect
of the resin of Gymnosperma glutinosum on the growth of A.
castellanii tro