Emerg Infect DisEIDEmerging Infectious Diseases1080-60401080-6059Centers for Disease Control and Prevention18507933260031407-169110.3201/eid1406.071691Letters to the EditorCoxiella burnetii in Wild-caught Filth FliesCoxiella burnetii in Wild-caught Filth FliesNelderMark P.*LloydJohn E.LoftisAmanda D.1ReevesWill K.§Clemson University, Clemson, South Carolina, USAUniversity of Wyoming, Laramie, Wyoming, USACenters for Disease Control and Prevention, Atlanta, Georgia, USAUS Department of Agriculture, LaramieCurrent affiliation: Private practice, Laramie, Wyoming, USAAddress for correspondence: Will K. Reeves, Agricultural Research Service, Arthropod-Borne Animal Diseases Research Laboratory, College of Agriculture, US Department of Agriculture, Department 3354, 1000 E University Ave, Laramie, WY 82071-2000, USA; email: will.reeves@ars.usda.gov6200814610021004Keywords: BlowflyQ feverstable flyzoozoonoticsheep kedCoxiella burnetiiMelophagus ovinusletter

To the Editor: Coxiella burnetii, the agent of Q fever, is a bacterium and a potential agent of bioterrorism. The most frequent signs of infection in domestic animals are abortion and reduced fertility (1). Clinical signs of Q fever in humans vary from mild fevers to pneumonia, hepatitis, or death; atypical cases occur as other disorders, such as cholecystitis (1,2). Aerosols are the most common route of exposure, but oral transmission occurs (1).

Some flies feed on the feces, milk, carcasses, or blood of domestic animals that can be infected with C. burnetii. These flies regurgitate and defecate when feeding and are mechanical vectors of bacteria (3,4). Flies have been shown to harbor, mechanically transport, and even support the growth of C. burnetii (46). It is known that house flies (Musca domestica) are possible mechanical vectors of C. burnetii because this organism survived 32 days in house flies and viable bacteria were shed by flies for 15 days (4). There are no studies of C. burnetii in field-collected flies. To examine the prevalence of C. burnetii in field-collected flies, we tested flies from farms, forests, ranches, and zoos.

Flies that develop on animal dung, carcasses, feces, blood, or garbage are often called filth flies. Adult Calliphoridae, Hippoboscidae, Muscidae, and Sarcophagidae were collected from forests, zoos, ranches, and farms (Table). Flies were killed in 95% ethanol or by freezing. DNA was extracted from individual flies as described (7,8). A distilled water negative control was used for each extraction.

Flies from the United States and Dominica assayed for <italic>Coxiella burnetii</italic>, 2004–2007
SpeciesCollection site and collectorDate of collectionNo. positive for C. burnetii/no. collected
Calliphora vicina (Robineau-Desvoidy)Elephant dung, Greenville Zoo, Greenville County, SC, USA, by M.P. Nelder2006 Apr 180/1
Lucilia coeruleiviridis (Marquart)Trapped on carrion near Pickens, SC, USA, by K.D. Cobb and W.K. Reeves2004 Jul 31/12
L. coeruleiviridis (Marquart)Elephant dung, Greenville Zoo, Greenville County, SC, USA, by M.P. Nelder2006 Jul 180/13
L. coeruleiviridis (Marquart)Elephant dung, Greenville Zoo, Greenville County SC, USA, by M.P. Nelder2005 Aug 170/3
L. sericata (Meigen)Garbage bin, Greenville Zoo, Greenville County, SC, USA, by M.P. Nelder2005 Aug 173/18
Melophagus ovinus (Linnaeus)Sheep, Bozeman, Gallatin County, MT, USA, by J.E. Lloyd2007 Jun 270/154
Musca domestica (Linnaeus)Cow, Springfield Estate, St. Paul Parish, Dominica, by W.K. Reeves2005 May 180/6
M. domesticaGoat pens, petting exibit, Greenville Zoo, Greenville County, SC, USA, by W.K. Reeves and M.P. Nelder2005 Aug 10/5
M. domesticaColobus spp. monkey dung; Greenville Zoo, Greenville County, SC, USA, by M.P. Nelder2005 Oct 190/6
Ravinia stimulansLion dung, Greenville Zoo, Greenville County, SC, USA, by M.P. Nelder2005 Oct 160/10
Ravinia new sp.Lion dung, Greenville Zoo, Greenville County, SC, USA, by M.P. Nelder2005 Oct 170/11
Stomoxys calcitrans (Linnaeus)Goat pens, petting exibit, Greenville Zoo, Greenville County, SC, USA, by W.K. Reeves and M.P. Nelder2005 Aug 10/20
S. calcitransFly trap, Greenville Zoo, Greenville County, SC, USA, by M.P. Nelder2006 Apr 280/20
S. calcitransCow, Riverbanks Zoo, Richland County, SC, USA, by M.P. Nelder2006 Apr 50/17
S. calcitransCow, goats, horses, and llama, Riverbanks Zoo, Richland County, SC, USA, by M.P. Nelder2006 Apr 60/12
S. calcitransCattle and elk hay barn, Sybille Canyon, Albany County, WY, USA, by W. Yarnell2007 July 14–Aug 31/55

Individual DNA samples were tested, in duplicate, with a previously described TaqMan assay with a lower limit of detection of 1 C. burnetii organism (8). Positive and negative controls were used for all assays. Positive flies were verified by PCR and sequencing of 16S rRNA gene as described (9). Vouchers for each insect species were deposited in the Clemson University Arthropod Collection (Clemson, SC, USA), the University of Georgia Museum of Natural History (Athens, GA, USA), or the University of Wyoming Insect Collection (Laramie, WY, USA).

Five of 363 flies were positive for C. burnetii DNA (Table). These flies included Stomoxys calcitrans, in which the adults feed on animal and human blood, and the blowflies Lucilia coeruleiviridis and L. sericata. C. burnetii–positive flies were obtained from carrion (1/12, 8.3%), a garbage bin of elephant feces (3/18, 16.7%), and a barn at a ranch (1/55, 1.8%). We sequenced 1,100 bp of the 16S rRNA gene from select DNA extracts, which were 99% identical with that of C. burnetii strain NC 002971.

We detected DNA from C. burnetii in flies from a zoo, a ranch, and carrion in a forest. Laboratory data on house flies, which shed live C. burnetii for 15 days after exposure, suggest that related flies (e.g., S. calcitrans and Lucilia spp.) might also harbor viable C. burnetii. On the basis of our field data, S. calcitrans and Lucilia spp. should be studied as mechanical vectors of C. burnetii. Unlike many enteric bacteria, which require large inocula to cause disease, C. burnetii can be infectious at the level of 1 bacterium (10). If flies transmit C. burnetii, they pose an additional threat to human and animal health.

The role of the sheep ked (Melophagus ovinus) in maintenance or transmission of C. burnetii is unknown. This fly is an obligate ectoparasite of sheep. It feeds on sheep blood, and feces from sheep keds can accumulate in the wool of sheep. Testing of sheep keds from infected sheep would help understand whether keds play a role in the epidemiology of C. burnetii.

Suggested citation for this article: Nelder MP, Lloyd JE, Loftis AD, Reeves WK. Coxiella burnetii in wild-caught filth flies [letter]. Emerg Infect Dis [serial on the Internet]. 2008 Jun [date cited]. Available from http://www.cdc.gov/EID/content/14/6/1002.htm

Acknowledgments

We thank A. Fabian, C. Kato, and R. Priestly for laboratory assistance; K.D. Cobb, G. Johnston, and W. Yarnell for field collections; J. Andre for research permits; T. Kreeger for access to his facility; R. Massung for positive-control DNA; and G. Dahlem for identifying Ravinia spp.

ReferencesWoldehiwet Z Q fever (coxiellosis): epidemiology and pathogenesis.Res Vet Sci 2004;77:93100 10.1016/j.rvsc.2003.09.00115196898Hartzell JD, Peng SW, Wood-Morris RN, Sarmiento DM, Collen JF, Robben PM, Atypical Q fever in US soldiers.Emerg Infect Dis 2007;13:1247917953104Nayduch D, Noblet GP, Stutzenberger FJ Vector potential of houseflies for the bacterium Aeromonas caviae.Med Vet Entomol 2002;16:1938 10.1046/j.1365-2915.2002.00363.x12109714Hucko M The role of the house fly (Musca domestica L.) in the transmission of Coxiella burnnetii.Folia Parasitol (Praha) 1984;31:177816745799Dhanda V, Padbidri VS, Mourya DT Multiplication of Coxiella burnetii in certain mosquitoes. In: Proceedings of the Symposium on Vectors and Vector-borne Diseases, Puri, Orissa, India. 1982 National Academy of Vector Borne Diseases. p. 69–73.Mourya DT, Padbidri VS, Dhanda V Mosquito inoculation technique for the diagnosis of Q fever employing an animal model.Indian J Med Res 1983;78:20146654408Kato CY, Mayer RT An improved, high-throughput method for detection of bluetongue virus RNA in Culicoides midges utilizing infrared-dye-labeled primers for reverse transcriptase PCR.J Virol Methods 2007;140:1407 10.1016/j.jviromet.2006.11.00917187871Loftis AD, Reeves WK, Szumlas DE, Abbassy MM, Helmy IM, Moriarity JR, Surveillance of Egyptian fleas for agents of public health significance: Anaplasma, Bartonella, Coxiella, Ehrlichia, Rickettsia, and Yersinia pestis.Am J Trop Med Hyg 2006;75:41816837707Reeves WK Molecular genetic evidence for a novel bacterial endosymbiont of Icosta americana (Diptera: Hippoboscidae).Entomol News 2005;116:2635Hatchette TF, Hudson RC, Schlech WF, Campbell NA, Hatchette JE, Ratnam S, Goat-associated Q fever: a new disease in Newfoundland.Emerg Infect Dis 2001;7:413911384518