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Operational mosquito and vector-borne diseases surveillance at Incirlik Air Base, Turkey.

Vector-borne and zoonotic diseases pose a threat to active military personnel both overseas and in the United States. Deaths among service members from vectorborne diseases have dropped in recent decades, but morbidity from vector-borne diseases and injuries from stinging arthropods remain a real threat. (1-4) The US Air Force (USAF) maintains arthropod identification and pathogen surveillance capability at the USAF School of Aerospace Medicine at Wright-Patterson Air Force Base, OH, through the Epidemiology Consult Service. Entomology services include the capability to identify both wide range of arthropods from around the world and pathogen surveillance including the detection of several arboviruses, Bartonella spp., Leishmania spp., Plasmodium spp., and Rickettsia spp. (5-10) Many of these identifications and pathogen screening services are used for both threat assessments and as part of standard pest management operations.

Long-term data on both vectors and associated pathogens are maintained for later review and determination of epidemiologic patterns. With the ongoing conflicts in the Middle East, analysis of trends in pathogen surveillance continues to be relevant. Several mosquito-borne arboviruses including West Nile virus (WNV) and Sindbis virus (SINV) were historically reported from both military members and local mosquitoes in the region. (11,12) We reviewed the arbovirus surveillance data for Incirlik Air Base (AB) in Turkey from 2011-2016 with a discussion of 2 major viruses.

Incirlik AB is in a strategically relevant location. It is located in far southern Turkey relatively near Syria and other Middle Eastern countries, including Iraq, Lebanon, and Israel. Surveillance is relevant for ongoing force health protection. In addition, pathogens detected in Incirlik could be useful in understanding viruses circulating in neighboring countries. Several mosquitoborne viruses such as West Nile and Sindbis are endemic in Turkey. (13,14)


The US Air Force currently maintains several military bases in the Middle East, but Incirlik AB, which opened in February 1955, has one of the longest established records. Most bases conduct vector and pathogen surveillance programs. Mosquitoes on many of these bases breed in water retention ponds or subsurface catch basins below parking lots or streets. Mosquitoes were trapped on the air base using the solid-state Army miniature traps with dry ice. Traps were set out seasonally from April through October and run on a weekly basis. Mosquito fogging typically occurred 3 days a week. Two traps were used, both near the perimeter fence, one near the sewage treatment plant, and one near base quarters. Mosquitoes were killed by freezing and shipped dry to the USAF School of Aerospace Medicine for identification and pathogen detection. Mosquitoes were morphologically identified using regional keys. (15,16) Voucher specimens were deposited in the Ohio State University Museum of Biological Diversity, Columbus, OH, or the Walter Reed Biosystematics Unit, Silver Spring, MD.

After identification, mosquitoes were pooled by species and tested for arboviruses. Pools ranged from 1-25 mosquitoes depending on submission numbers. Most mosquitoes were tested with a RAMP* WNV test and the VecTOR Test Systems (Thousand Oaks, California) Alphavirus panel that detects chikungunya (CHIK), equine encephalitis, Mayaro, SINV, Venezuelan equine encephalitis, and Western equine encephalitis viruses. These rapid assays which had been designed for field deployment and rapid screening of mosquitoes (17) were among the tools used from 2011-2016 by the USAF School of Aerospace Medicine. Both the VecTOR Test Systems and RAMP assays were performed in accordance with manufacturers" guidelines. We occasionally used commercially available inactivated West Nile and chikungunya virus antigens as a positive control to validate the assay sensitivity as suggested. (18) As a further validation, we verified 2 of the positive pools by reverse transcription polymerase chain reaction (RT-PCR). West Nile virus positive pools were verified using the protocols by Lanciotti et al. (19) The PCR products were sequenced and compared to sequences in GenBank using the BLAST ([dagger]) program.

There were 6 pools of Culex perexiguus Theobald or Cx. pipiens L. that tested positive for arboviruses as shown in the Table. There were 4 pools positive for WNV: one each on October 2 and 9, 2013; August 29, 2014; and September 16, 2015. We validated the October 9, 2013 results with both the RAMP and VecTOR Test Systems WNV assays. The RAMP scores ranged from 150 to the maximum limit of 740. Both pools tested by RT-PCR were positive with cycle threshold scores of 18.14 and 21.04 respectively which was less than the viral culture positive controls. The 919 base-pair sequence of WNV envelope protein was a 100% match to WNV isolate Spain/2010/H-1b (GenBank # JF719069).

In addition, 2 pools of combined Cx. perexiguus and Cx. pipiens from June 18 and 25, 2014, tested strongly positive for an Alphavirus on the VecTOR Test System assay. This assay is designed to detect specific viruses, and Sindbis is the detectable virus in the region.13


We detected West Nile and Sindbis viruses in pools of Culex spp. from Incirlik AB. The WNV was in the Western Mediterranean WNV subtype in the WNV lineage 1 based on sequence similarity. While often underappreciated, both viruses are a true threat to the military. Petersen et al (20) estimated that WNV caused hundreds of thousands of infections in the United States annually from 1999 to 2010. Further, WNV was the leading cause of vector-borne disease death in USAF-associated individuals since the 1970s. (2) Sindbis virus infection causes severe arthritis, fever, and, in rare cases, encephalitis. (21)

West Nile virus was detected in a total of 6 collections made in 2013, 2014, and 2015. This indicates that the virus circulates at some regularity and could pose a threat to both active duty, contractors, civilian personnel, and dependents. We separated mosquito collection data between sewage or waste water treatment plants and residential areas and detected infected mosquitoes in both areas. This virus requires avian hosts for the natural enzootic cycle but mosquitoes can be transovarially infected. (22) We could not determine the route of infection in mosquitoes from Incirlik AB.

Incirlik AB is close to Syria and other conflict zones in the Middle East. Detection of 2 potentially serious arbovirus threats on a military base in southern Turkey could raise concerns for similar disease threats in neighboring countries. This is especially true in conflict zones with little to no infrastructure. Infections by either West Nile or Sindbis virus often lead to mild disease but can lead to fatal or debilitating infections and might result in costly medical evacuations.

In summary, we reviewed 6 years (2011-2016) of WNV and SINV detection in mosquitoes from Incirlik AB. Culex perexiguus was the most commonly associated mosquito with WNV from the base. It is a member of the Cx. univittatus Theobald complex and several of these have previously been associated with both WNV and SINV with evidence of vertical transmission of WNV. (23) Our surveillance data suggests that there are repeated and predictable threats from WNV and possibly SINV on the base.


We thank the Public Health and Civil Engineering personnel at Incirlik AB for mosquito surveillance and timely submission of samples. We also thank the current and former Entomology staff from the US Air Force School of Aerospace Medicine.

This research was partially funded by the Global Emerging Infections Surveillance and Response System.

This article was cleared by the Defense Office of Prepublication and Security Review, Security Review# 17-S-1077.


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Will K. Reeves, PhD, MS

Myrna M. Miller, DVM, PhD

Orhan Bayik, PHS

Maj Leah Chapman, USAF, MS


When this study was conducted, Dr Reeves was the USAF consult entomologist at the the USAF School of Aerospace Medicine. He currently is a regulatory analyst at the US Department of Agriculture Animal and Plant Health Inspection Service, Biotechnology Regulatory Service, Fort Collins, Colorado.

Dr Miller is the chief virologist at the Wyoming State Veterinary Laboratory, Laramie, Wyoming.

Mr Bayik is the Public Health Specialist and Food Safety and Medical Entomology Program Manager at the Incirlik Air Base, Turkey.

When this study was conducted, Maj Chapman was a Public Health instructor at the USAF School of Aerospace Medicine. She currently is the Assistant Contingency Liaison Officer at the Armed Forces Pest Management Board, Silver Spring, Maryland.

* RAMP (rapid analyte measurement platform) is a registered trademark of Response Biomedical, Vancouver, Canada.

([dagger]) Basic Local Alignment Search Tool; National Center for Biotechnology Information, Bethesda, MD.
Mosquito surveillance data from Incirlik Air Base, Turkey, from
2011-2016 with WNV virus and Sindbis virus screening data.

Locality name      Collection Dates

Housing            2 June -19 August 2011

Housing            27 April-14 September 2012

Running Track      1 June 2012

Housing            18 April- 20 November 2013

Sewage Treatment   18 April- 20 November 2013

Flight Line        8 April -6 November 2013
Housing            16 May-29 August 2014

Dining Facility    2 July 2014

Sewage Treatment   23 April- 29 August 2014

Housing            6 May-23 September 2015

Sewage Treatment   1 May-16 September 2015

Housing            12 April-12 August 2016

Sewage Treatment   31 March-16 October 2016

Locality name      Species * and Number

Housing            Culex perexiguus (95)
                   Culex pipiens (23)
                   Aedes caspius (1)
                   Culiseta longiareolata (7)
Housing            Culex perexiguus (133)
                   Cules pipiens (388)
                   Culex mimeticus (7)
                   Aedes caspius (7)
                   Culiseta longiareolata (6)
Running Track      Culex perexiguus (5)
                   Culex pipiens (13)
Housing            Culex perexiguus (73)
                   Culex pipiens (339)
                   Culex sinaiticus (148)
                   Aedes caspius (17)
                   Culiseta longiareolata (7)
Sewage Treatment   Culex perexiguus (821)
Plant              Culex pipiens (252)
                   Culex sinaiticus (301)
                   Aedes caspius (32)
Flight Line        Culex pipiens (8)
Housing            Anopheles claviger (1)
                   Culex perexiguus (658)
                   Culex pipiens (153)
                   Culiseta longiareolata (1)
Dining Facility    Culex perexiguus (1)
                   Culex pipiens (3)
Sewage Treatment   Culex perexiguus (327)
plant              Culex pipiens (90)
                   Aedes capensis (9)

Housing            Culex perexiguus (331)
                   Culex pipiens (280)
                   Aedes caspius (11)
                   Culiseta longiareolata (14)
Sewage Treatment   Culex perexiguus (831)
Plant              Culex pipiens (198)
                   Aedes caspius (2)
                   Culiseta longiareolata (3)
Housing            Culex perexiguus (11)
                   Culex pipiens (33)
                   Culiseta longiareolata (3)
Sewage Treatment   Culex perexiguus (225)
Plant              Culex pipiens (182)
                   Aedes capensis (3)

Locality name      Arbovirus Test Results

Housing            WNV/Alphavirus Negative

Housing            WNV/Alphavirus Negative

Running Track      WNV/Alphavirus Negative

Housing            One pool of 25 Culex
                   perexiguus from 9 October 2013
                   was positive for WNV virus.

Sewage Treatment   One pool of 25 Culex
Plant              perexiguus from 2 October
                   2013 was
                   positive for WNV virus.
Flight Line        WNV/Alphavirus Negative
Housing            WNV/Alphavirus Negative

Dining Facility    WNV/Alphavirus Negative

Sewage Treatment   Two pools of combined Culex
plant              spp. (25 each) from 18 and 25
                   June 2014 tested positive for
                   Alphavirus. One pool of 25
                   Culex perexiguus from 29
                   August 2014 was positive for
                   WNV virus.

Housing            One pool of 25 Culex
                   perexiguus from 16 September
                   2015 was positive for WNV
Sewage Treatment   WNV/Alphavirus Negative

Housing            WNV/Alphavirus Negative

Sewage Treatment   WNV/Alphavirus Negative

* Scientific species identification:
Aedes capensis Edwards
Aedes caspius (Pallas)

Culiseta longiareolata Macquart
Culex mimeticus Noe

Culex perexiguus Theobald
Cules pipiens L.

Culex sinaiticus Kirkpatrick
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Article Details
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Author:Reeves, Will K.; Miller, Myrna M.; Bayik, Orhan; Chapman, Leah
Publication:U.S. Army Medical Department Journal
Article Type:Report
Geographic Code:7TURK
Date:Jan 1, 2017
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