CDC Dengue Branch Publication List for 2009 Munoz-Jordan, J: Tomashek KM, Rivera A, Muoz-Jordan JL, Hunsperger E, Santiago

L, Padro O, Garcia E, Sun W: Description of a large island-wide outbreak of dengue in Puerto Rico, 2007. Am J Trop Med Hyg 2009, 81:467-474. Muoz-Jordan JL, Collins CS, Vergne E, Santiago GA, Petersen L, Sun W, Linnen JM: Highly sensitive detection of dengue virus nucleic acid in samples from clinically ill patients. J Clin Microbiol 2009, 47:927-931. Hunsperger EA, McElroy KL, Bessoff K, Colon C, Barrera R, Muoz-Jordan JL: West Nile virus from blood donors, vertebrates, and mosquitoes, Puerto Rico, 2007. Emerg Infect Dis 2009, 15:1298-1300. de Thoisy B, Lacoste V, Germain A, Muoz-Jordan JL, Colon C, Mauffrey JF, Delaval M, Catzeflis F, Kazanji M, Matheus S, et al.: Dengue infection in neotropical forest mammals. Vector Borne Zoonotic Dis 2009, 9:157-170. Vasilakis N, Durbin AP, da Rosa AP, Muoz-Jordan JL, Tesh RB, Weaver SC: Antigenic Relationships between Sylvatic and Endemic Dengue Viruses. Am J Trop Med Hyg 2008, 79:128-132. Rondini S, Pingle MR, Das S, Tesh R, Rundell MS, Hom J, Stramer S, Turner K, Rossmann SN, Lanciotti R, Muoz-Jordan JL et al.: Development of multiplex PCR-ligase detection reaction assay for detection of West Nile virus. J Clin Microbiol 2008, 46:2269-2279. Ramos MM, Mohammed H, Zielinski-Gutierrez E, Hayden MH, Lopez JL, Fournier M, Trujillo AR, Burton R, Brunkard JM, Anaya-Lopez L, Muoz-Jordan JL et al.: Epidemic Dengue and Dengue Hemorrhagic Fever at the Texas-Mexico Border: Results of a Household-based Seroepidemiologic Survey, December 2005. Am J Trop Med Hyg 2008, 78:364-369. Ramos MM, Arguello DF, Luxemburger C, Quinones L, Muoz-Jordan JL, Beatty M, Lang J, Tomashek KM: Epidemiological and clinical observations on patients with dengue in Puerto Rico: results from the first year of enhanced surveillance--June 2005-May 2006. Am J Trop Med Hyg 2008, 79:123-127. Mohammed H, Linnen JM, Muoz-Jordan JL, Tomashek K, Foster G, Broulik AS, Petersen L, Stramer SL: Dengue virus in blood donations, Puerto Rico, 2005. Transfusion 2008, 48:13481354. Das S, Pingle MR, Muoz-Jordan JL, Rundell MS, Rondini S, Granger K, Chang GJ, Kelly E, Spier EG, Larone D, et al.: Detection and serotyping of dengue virus in serum samples by

multiplex reverse transcriptase PCR-ligase detection reaction assay. J Clin Microbiol 2008, 46:3276-3284. Barrera R, Hunsperger E, Muoz-Jordan JL, Amador M, Diaz A, Smith J, Bessoff K, Beltran M, Vergne E, Verduin M, et al.: First isolation of West Nile virus in the Caribbean. Am J Trop Med Hyg 2008, 78:666-668.

Abstract
A sentinel chicken program for West Nile virus (WNV) surveillance was initiated in July 2006 in eastern Puerto Rico, yielding the first seroconversions on June 4, 2007. WNV was isolated from sentinel chicken serum and mosquito pools (Culex nigripalpus, Culex bahamensis) for the first time in Tropical America. Preliminary sequence analysis of the prM and E genes revealed a 1amino acid difference (V159A) between the Puerto Rican 2007 and the NY99. This mutation has been observed in the current dominant clade circulating in the United States. Sentinel chicken surveillance was a useful tool for the detection of West Nile virus in the tropics. WNV has been spreading southward into the Caribbean Basin and Latin America since 2001 when the first human case was reported from the Cayman Islands.1 Serological evidence of WNV transmission has been accumulating, and cross-reactive WNV antibodies have been detected in humans from Mexico, The Bahamas, and Cuba, in horses from Guadalupe, Mexico, Central America, Cuba, Puerto Rico, Colombia, and Venezuela, and in resident birds from Ja-maica, Dominican Republic, Cuba, Puerto Rico, and Venezu-ela.26 In November 2006, 4 serologically confirmed human WNV encephalitis cases were reported by Argentina. 7 WNV may be following the same patterns of southward dissemination as other arboviruses into the Caribbean and Central and South America via migratory birds.8 In spite of growing evidence for the presence of WNV in tropical America, no outbreaks have yet been reported from the region. It is also interesting to note that high bird mortality, as observed in the Nearctic, has not been reported. WNV has been isolated in tropical America from a dead raven in Tabasco, Mexico (18 N lat.),9 and in subtropical Argentina (2349 S lat.) from dead horses,10 but no isolates have yet been reported from other areas in Central and South America or the Caribbean. One explanation for the paucity in WNV detection in tropical America may be the lack of active WNV surveillance. One would hypothesize that the likelihood of identifying or isolating WNV would be maximized in the Caribbean Basin, given its location in the flight paths of infected migratory birds. Another plausible explanation is that WNV has not been able to establish persistent enzootic foci in the region, so that reported serological evidence may be caused by frequent but unsuccessful virus re-introductions. Here, we report the first WNV isolation in the Caribbean, specifically from sentinel chicken serum and mosquitoes in natural and rural areas of eastern Puerto Rico. To investigate the presence of WNV in Puerto Rico, a sentinel chicken program was established in July 2006 in Ceiba and Naguabo municipalities, near the place where a resident bird was previously found with WNV antibodies 3 and close to the area where the Puerto Rico passive surveillance system detected 3 asymptomatic horses with WNV antibodies in 2004.11 The research protocol was approved by CDCs Animal Care and Use Committee (06-012). Sixty sentinel chickens were pretested for flavivirus reactive antibodies and placed in 12 pens, each housing 5 chickens. The 12 pens were placed in 8 natural habitats (2 herbaceous wetlands, 2 mangrove forests, 2 deciduous forests, and 2 evergreen forests) and in 4 nearby human populated areas (2 rural and 2 urban areas). Blood samples were taken from all sentinel chickens every 2 weeks and tested by a chicken-specific MAC-ELISA 12; 1,247 blood samples were analyzed before initial seroconversion was detected (JulyJune 2006). The first seroconversion of 7 chickens (12%) was detected in the sentinel chickens in 4 pens from natural areas (1 wet-land, 1 mangrove forest, and 2 evergreen forests) in June 4, 2007. Following initial seroconversion, bleeding and sentinel replacement were performed weekly.

Seroconversion increased to 40% on June 11, 2007, and was detected in chickens from 11 out of the 12 pens, covering all types of habitats including rural and urban areas. Chickens continued to sero-convert at high rates (max. 45%) throughout the rest of June and July, but the rate sharply declined during August 2007 (2%). Seroconversion has continued through October 2007 indicating a sustained low level of transmission (26%). Plaque reduction neutralization tests (PRNT90)13 against WNV and Saint Louis encephalitis virus (SLE) were used to confirm that the infecting virus was WNV (Table 1). Serum samples were heat inactivated at 56C for 30 minutes and serially diluted 2-fold in phosphate-buffered saline (PBS) with 30% heat-inactivated fetal bovine serum. One hundred microliters of 1:20 diluted serum was mixed with an equal volume of diluents containing ChimeraVax WNV or SLE virus and analyzed in duplicate (Acambis, Cambridge, MA). One hundred microliters of the serumvirus suspension was used to inoculate a confluent monolayer of Vero cells in 12-well plates and incubated at room temperature for 1 hour. Two milliliters of medium containing 10% M199 without phenol red, 1% essential amino acids, 1% vitamins, 1% glutamine, 5% inactivated FBS, 0.4% gentamicin, 4% sodium bicarbonate, and 0.6% agarose was added to each well and placed in a 37C 10% CO2 incubator for 4 days for WNV and 5 days for SLE virus. Plaques were stained by adding 500 L of PBS containing 3.2% neutral red to each well at 3 or 4 days post-infection for WNV and SLE virus, respectively. Plaques were counted 24 hours later, and endpoint titers were expressed as the reciprocal of serum dilutions yielding 90% reduction in the number of plaques. Controls included virus only and virus with normal serum.13 Chicken serum specimens that were collected a week prior to seroconversion were tested by RTPCR to attempt virus isolation from these samples. 14 This led to positive identification of WNV RNA in 1 of the sentinel chickens and subsequently WNV isolation from this chicken serum in Vero cells. Mosquito captures were performed around the positive pen areas using CDC miniature light/CO2 traps and CDC gravid traps. Captured mosquitoes were preserved on dry ice, sorted in pools (50 females per pool), and stored at -70C until they were tested by RT-PCR. 15 The following mosquito species were initially found infected with WNV: Culex nigripalpus (51 positive out of 101 pools tested; 50.5%), Culex bahamensis (3/8; 37.5%), and Culex quinquefasciatus (4/46; 8.7%). WNV isolations were made in both C6/36 and Vero cells16: 4 from Cx. nigripalpus pools and 1 Cx. bahamaensis. Virus isolations were confirmed by RT-PCR and by specific immunostaining using West Nile/Kunjun 393 monoclonal antibody (CDC, catalog no. m28955A) and FITC-conjugated anti-mouse IgG (KPL, Inc., Gaithersburg, MD) in the presence of 4,6 di-amino-2-phenylindole, DAPI (Sigma, St. Louis, MO) according to previously published procedures in both cell lines.16 Preliminary sequence analysis of the prM and E genes (Gen-Bank library accession number: EU394703) revealed one amino acid difference (V159A) between Puerto Rican 2007 and NY99. This mutation has been observed in the current dominant clade circulating in the United States. 17 No clinical cases of humans or equines had been reported by the end of July. Enhanced surveillance for human WNV infection has been implemented. These results demonstrate that WNV is actively circulating in Puerto Rico and that sentinel chickens and mosquito surveillance were useful tools for detecting ongoing WNV transmission in areas with pre-existing serologic evidence from a resident bird and horses. In spite of the efforts that were made to capture mosquitoes (JuneJuly 2004) following the recovery of IgG antibodies in a resident bird (February 2004) and 3 horses (May 2004), no mosquito pool was found positive by TaqMan RT-PCR that year (CDC, unpublished data). It is likely that detection of WNV IgG antibodies was not an adequate indicator of ongoing or recent WNV transmission. However, WNV surveillance on resident birds and horses using IgG antibodies was useful to identify areas of previous virus transmission, and further surveillance using IgM antibodies in sentinel chickens was useful to detect active transmission at those sites. Although a passive WNV surveillance system on the islands had been in place since 2002 (Puerto Rico Department of Health and CDCs Dengue Branch; unpublished data), only 3 horses out of 4,370 specimens (from dead birds, freeranging and domestic fowl, domestic porcine, equine, canine, caged primates, and humans) were found infected with WNV-reactive antibodies. Thus, we recommend sustained WNV surveillance with sentinel chickens in areas where the presence of this virus is suspected. Furthermore,

human WNV surveillance in Puerto Rico, where dengue is endemic, will require careful interpretation of serologic tests and assiduous efforts to isolate the infecting virus. Our preliminary genetic analysis of the WNV Puerto Rico isolate from a sentinel chicken resulted in a 99.8% homology with NY99 strain, and further studies are being conducted to determine its pathogenesis.

Link: http://www.ajtmh.org/content/78/4/666.full XXXBarrera R, Amador M, Diaz A, Smith J, Muoz-Jordan JL, Rosario Y: Unusual productivity of Aedes aegypti in septic tanks and its implications for dengue control. Med Vet Entomol 2008, 22:62-69. Tomashek, K: (I published 6 peer-reviewed manuscripts (three as member of AAP COFN -- the LAST) and 1 MMWR in 2009 -- up through Sept; I was first or second author for 2 of the manuscripts. I have six additional manuscripts in review at peer-reviewed journals; 5 were just accepted for publication and are in press (will be in print in 2009 or early 2010). We should have a MMWR for Key West and a peer-reviewed journal article into review by end of 2009.) Peer Reviewed Journal Articles XXXPouliot SH, Xiong X, Harville E, Paz-Soldan V, Tomashek KM, Breart G, Buekens P. Maternal Dengue and Pregnancy Outcomes: A Systematic Review. (Obstetrical & Gynecological Survey, in press.) XXXGregory CJ, Llata E, Stine N, Gould C, Santiago LM, Vazquez GJ, Robledo IE, Srinivasan A, Goering RV, Tomashek KM. Outbreak of Carbapenem-Resistant Klebsiella pneumoniae in Puerto Rico Associated with a Novel Carbapenemase Variant. (Infection Control and Hospital Epidemiology, in press) Gregory CJ. Santiago LM, Arguello DF, Muoz-Jordan JL, Hunsperger E, Tomashek KM. Clinical and Laboratory Features that Differentiate Dengue from Other Febrile Illnesses in an Endemic Area Puerto Rico, 2007-2008. (American Journal of Tropical Medicine and Hygiene, in review.)

Abstract.
Dengue infection can be challenging to diagnose early in the course of infection before severe manifestations develop, but early diagnosis can improve patient outcomes and promote timely public health interventions. We developed age-based predictive models generated from 2 years of data from an enhanced dengue surveillance system in Puerto Rico. These models were internally validated and were able to differentiate dengue infection from other acute febrile illnesses with moderate accuracy. The accuracy of the models was greater than either the current World Health Organization case definition for dengue fever or a proposed modification to this definition, while requiring the collection of fewer data. In young children, thrombocytopenia and the absence of cough were associated with dengue infection; for adults, rash, leucopenia, and the absence of sore throat were associated with dengue infection; in all age groups, retroorbital pain was associated with dengue infection. Link: http://www.ajtmh.org/content/82/5/922.full

XXXLeiss J., Giles D., Sullivan K.M., Mathews R., Sentelle G., Tomashek KM. Are U.S. maternally linked birth records biased with respect to linkages among marginal population groups? (Annals of Epidemiology, in press). XXXNannini A, Lazar J, Berg C, Tomashek KM, Barfield WD, Barger M, Cabral H, Kotelchuck M. Timing of hospital visits for assault during the pregnancy-associated period. (Journal of Women's Health, in press). Mohammed H, Ramos MM, Rivera A, Garcia E, Sun W, Tomashek KM. Travel-Associated Dengue Infections in the United States, 1996-2005. (The Journal of Travel Medicine, in press). Tomashek KM, Rivera A, Santiago L, Muoz-Jordan JL, Hunsperger E, Padro O, Garcia E, Sun W. Description of a large island-wide outbreak in Puerto Rico, 2007. American Journal of Tropical Medicine and Hygiene 2009; 81(3):467-74. Shapiro-Mendoza CK, Kimball M, Tomashek KM, Anderson R, Blanding S. US Infant Mortality Trends Attributable to Accidental Sleep-related Suffocation and Strangulation Deaths from 1984 through 2004: Are Rates Increasing? Pediatrics 2009; 123(2):533-39. Ramos MM, Tomashek KM, Arguello DF, Luxemburger C, Quiones L, Lang J, Muoz-Jordan JL. Early clinical features of dengue infection in Puerto Rico. Transactions of the Royal Society of Tropical Medicine and Hygiene 2009 Sep; 103(9):878-84. Peer Reviewed Journal Articles as CDC Liaison to AAPs Committee on Fetus and Newborn Batton DG; Committee on Fetus and Newborn. Clinical report--Antenatal counseling regarding resuscitation at an extremely low gestational age. Pediatrics. 2009 Jul;124(1):422-7. Committee on Fetus and Newborn, Wallman C. Advanced practice in neonatal nursing. Pediatrics. 2009 Jun;123(6):1606-7. Bull MJ, Engle WA; Committee on Injury, Violence, and Poison Prevention and Committee on Fetus and Newborn; American Academy of Pediatrics. Safe transportation of preterm and low birth weight infants at hospital discharge. Pediatrics. 2009 May;123(5):1424-9. Morbidity and Mortality Weekly Review (MMWR) Articles Lledo W, Hernandez M, Lopez E, Molinari OL, Soto RQ, Hernandez E, Santiago N, Flores M, Vazquez GJ, Robledo IE, Garcia-Rivera E, Cortes A, Ramos M, Goering R, Srinivasan A, Gould C, Stine N, Bell M, Anderson K, Kitchel B, Wong B, Rasheed JK, Patel J, Tomashek K, Llata E, Gregory CJ. Guidance for Control of Infections With Carbapenem-Resistant or Carbapenemase-Producing Enterobacteriaceae in Acute Care Facilities. MMWR 2009; 58:256260 (and reprint in JAMA 2009; 301(19):1980-82). Book Chapter Tomashek K. Dengue Fever and Dengue Hemorrhagic Fever. In: Centers for Disease Control and Prevention. CDC Health Information for International Travel 2010. Atlanta: U.S. Department of Health and Human Services, Public Health Service, 2009.

Barrera, R: Mackay AJ, Amador M, Diaz A, Smith Josh and Barrera R. (2009). Dynamics of Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae) in septic tanks in Southern Puerto Rico. Journal of the American Mosquito Control Association. Barrera R. 2009. Geospatial studies of the ecology and control of mosquito vectors and dengue transmission. In: Approaches and Current Themes in Entomology. Arrivillaga J, El Souki M, Herrera B. eds., Editorial Astrodata, Caracas, Venezuela. (in Spanish) Hunsperger EA, McElroy KL, Bessoff K, Coln C, Barrera R, Muoz-Jordn JL. 2009. West Nile Virus Presence in Human, Equine, Avian and Mosquito Species in Puerto Rico, 2007. Emerging Infectious Diseases 15:1298-1300. Barrera R. 2009. Simplified Aedes aegyptis pupal-surveys for entomological surveillance and dengue control. American Journal of Tropical Medicine and Hygiene 81: 100-107. Smith J, Amador M, Barrera R. 2009. Seasonal and Habitat Effects on Dengue and West Nile Virus Vectors in San Juan, Puerto Rico. Journal of the American Mosquito Control Association 25: 38-46. Barrera R, Amador M, Diaz A. Joshua Smith, Munoz-Jordan JL, Rosario Y. 2008. Unusual productivity of Aedes aegypti in septic tanks and its implications for dengue control. Medical and Veterinary Entomology 22:62-69 Barrera R, Hunsperger E, Muoz-Jordn, Amador M, Diaz A, Smith J, Bessoff K, Beltran M, Vergne E, Verduin M, Sun W. 2008. First isolation of West Nile virus in the Caribbean. Journal of Tropical Medicine and Hygiene 78:666-669 Johansson, M: Johansson, MA, F Dominici, & GE Glass. Local and global effects of climate on dengue transmission in Puerto Rico. PLoS Neglected Tropical Diseases. 3(2): e382 (2009). Reed, LM, MA Johansson, N Panella, R McLean, T Creekmore, R Puelle, & N Komar. Declining mortality in American crow (Corvus brachyrhynchos) following natural West Nile virus infection. Avian Diseases. 53: 458-461 (2009). Johansson, MA, DAT Cummings, & GE Glass. Multi-year Climate Variability and Dengue - El Nio Southern Oscillation, Weather, and Dengue Transmission in Puerto Rico: A Longitudinal Data Analysis. PLoS Medicine. (scheduled publication date: Nov 17, 2009) Hunsperger, E.:

Hunsperger EA, Roehrig J. (2009) Nocodazole delays viral entry into the brain following footpad inoculation with West Nile virus in mice. JNeuroVirol 15:211-218. Johnson B, Kosoy O, Hunsperger E, Beltran M, Delorey M, Guirakhoo F, Monath T. (2009) Evaluation of chimeric Japanese encephalitis and dengue viruses for use in diagnostic plaque reduction neutralization tests. Clin Vaccine Immunol 16(7):1052-1059. Hunsperger EA, McElroy K, Bessoff K, Colon C, Barrera R, Muoz-Jordan J. (2009) Isolation and analysis of West Nile virus from blood donors, vertebrate animals and mosquitoes in Puerto Rico, 2007. EID 15(8):1298-1300. Tomashek KM, Rivera A, Hunsperger E, Muoz-Jordan JL, Santiago L, Padr,O Garcia E, Sun W. (2009) Description of a Large Island-Wide Outbreak of Dengue in Puerto Rico, 2007. AJTMH 81(3):467-474. Hunsperger E, Yoksan S, Buchy P, Nguyen VC, Sekaran SD, Enria DA, Pelegrino JL, Vazquez S, Artsob H, Drebot M, Gubler D, , Halstead SB , Guzmn MG, Margolis HS, Nathanson CM, Rizzo NR, Bessoff KE, Kliks S, Peeling RW. (2009) Evaluation of commercially available dengue IgM tests by a WHO/PDVI Laboratory Network, EID 15(3):436-440. Currently in review from various journals: Phoutrides E, Jusino-Mendez, Prez-Medina T, Seda-Lozada R, Garcia-Negron M, Davila-Toro F, Carazo C, Hunsperger E. Effectiveness of animal surveillance during the reemergence of West Nile virus in Puerto Rico, 2007. In review EID. Komar N, Hunsperger E, Bessoff K, Amador M, Barrera R. Avian host of West Nile virus in Puerto Rico. In review AJSTMH 2009. Costin JM, Hrobowski YM, Rees CR, Fontaine KA, Holbrook MR, Hunsperger E, Garry RF, Isern S, Michael SF. A broadly active peptide inhibitor of flavivirus entry. Plos Neglected Diseases in review 2009. Mercado X, Rivera Y, Hunsperger E, Martnez I. Dengue Virus Type 2 Confers Cross Protection against West Nile Virus Induced Disease in Mice, In review AJSTMH, 2008. Mohammed, H: Travel-Associated Dengue Infections in the United States, 1996 to 2005; Hamish P. Mohammed, PhD, Mary M. Ramos, MD, Aidsa Rivera, MSc, Michael Johansson, PhD, Jorge L. Muoz-Jordan, PhD, Wellington Sun, MD and May M. Tomashek, MD; Journal of Travel Medicine 2009.