Progress 09/01/16 to 08/31/18
Outputs
Target Audience:Information obtained in this study couldbe helpful to public health workers and government agencies who are involved in preventing the spread of Zika virus. Agencies and departments such as the Florida Department of Health, Ministry of Health in Haiti, Centers for Disease Control wouldfind the information useful. Inaddition, data on the mosquitoes capable of transmitting the virus could help Mosquito control districts in Haiti and in Florida in thecontrol efforts. Changes/Problems:The problems of storage and shipping of mosquitoes from Haiti to the US. In addition, the high volume of mosquitoes that were caught that required expert identification significanly slowed projec progress. We will continue to identify the mosquitoes caught and are in the process of developing new proposals to help us strengthen entomology techncian support in mosquito identification in Haiti. What opportunities for training and professional development has the project provided?The project provided 2 graduate students with the skills in virus detection by RT-PCR, and virus isolation of Zika. In addition, a postdoctoral trainee also benefited in the work. How have the results been disseminated to communities of interest?Results have been presented at local andnational conferences. In addition, small meetings to provide status updates officials of theHaiti Ministry of Health have been conducted. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Objective 1. Determine the frequency of ZIKV in wild caught mosquitoes in households in Haiti. Major activities completed / experiments conducted. Mosquitoes were trapped are several locations in Haiti. The mosquito collection sites included 3 sites in Gressier (Mareshal, Merger and Lacolin) and 3 other sites in Leogane (Belval, Cira, Signueneu) in Leogane. Biogents sentinel traps and CDC light traps were set up at strategic locations. Captured mosquitoes were sorted out by species, sexed and stored at minus 80 degrees Centigrade freezer to await further analysis at University of Florida labs in Gainesville. Data collected. Mosquito species captured by location over time. Infection status of mosquitoes over different localities and over several months. Laboratory experiments: Species and sex specific mosquito pools were made and homogenized in refrigerated phosphate buffered saline (PBS), centrifuged and the supernatant aliquoted (a) into lysis buffer for immediate viral RNA (vRNA) extraction using a Qiagen QIAamp viral RNA mini kit (Qiagen, USA), and (b) PBS containing 15% trehalose for storage at -80°C for cryopreservation of virus particles for isolation attempts in cell cultures. If an original pool contained more than 20 mosquitoes, the pool was divided into sub-pools for homogenization.Extracted nucleic acids were subsequently screened by real-time (rt)RT-PCR for ZIKV vRNAs using published protocols. Some pools that yielded a positive result were subsequently inoculated onto Vero E6 cells and incubated at 37°C in 5% CO2 for up to 30 days for virus isolation attempts. Upon observation of virus-specific cytopathic effects (CPE) throughout 50% of the monolayer, spent media and scraped cells in spent media were collected and tested by molecular methods for ZIKV vRNAs. Additionally, the mosquito species (Ae. aegypti, Ae. albopictus, and Culex quinquefasciatus), and Ae. aegypti sex were confirmed in virus-positive pools by molecular methods using published protocols. Both the homogenate and the spent media were used for whole genome sequencing by Sanger sequencing methods as previously reported. Summary of results. A total of 49, 853 mosquitoes were captured in the period August 2016 to July 2017. Additionally mosquitos were collected between March 2018 to June 2018. In the 2016 collections, 10,688 were collected in Gressier and 15,083 were collected in Leogane. In 2018, 10, 658 mosquitoes were collected from 4 additional locations in Gressier (3 locations) and Loegane (1). Of these mosquitoes, 1235 were Aedes aegypti, 33 were Ae albopictus, 29 were Ae mediovittatus, 7143 were Culex quinquefasciatus, 1798 were Culex nigrapalpus. The rest of the mosquitoes were not identified but included Culex, Psophora and Orthopodomyia species. Zika infections in mosquito . Mosquitoes collected in 2016 to 2017 were tested for Zika virus infection. For zika virus detection, 296 pools of Aedes aegypti were homogezized and tested. Twenty eight pools were positive or Zika virus infection. Of these pools, 206 were female Aede aegypti and 90 were male Aedes aegypti. All the collection sites yielded Zika positive mosquitoes - Leogane 1 - 6 pools (9.2%); Leogane 2 - 7 pools, Leogane 3 - 7 pools, Gressier 1 - 35, Gressier 2 - 22 pools, Gressier 3 - 28 pools. Over a collection period of 2017, from January to May, Zika virus positive mosquito pools were detected from the collection sites with the highest positves in Feburary 2017 where 16.7% of the pools were positive. [January - 15.4% (4/26); February - 16.7% (7/42); March 3% (3/50); April - 2% (2/95); May 14.4% (12/83).
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
White SK, JA Lednicky, JG Morris, Jr., JC Dunford, and BA Okech. 2017. Detection of Chikungunya-, Dengue and Zika viruses in mosquitoes collected in Haiti, 2016. University of Florida Emerging Pathogens Institute Research Days, Gainesville, Florida.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
White SK, JA Lednicky, J Morris, Jr., B Okech, and J Dunford. 2017. Detection of Chikungunya-, Dengue and Zika viruses in mosquitoes collected in Haiti, 2016. Entomological Society of America Annual Conference, Denver, Colorado
- Type:
Journal Articles
Status:
Submitted
Year Published:
2018
Citation:
Sarah K. White, John A. Lednicky, Bernard A. Okech, J. Glenn Morris Jr
(2018. Evidence of vertical transmission and establishment of Zika virus in field-caught mosquitoes, Haiti 2016
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