LETTERS spp. isolates in US food animals (10). 4. Galimand M, Guiyoule A, Gerbaud G, Triatoma infestans Given these linkages, the transfer of an Rasoamanana B, Chanteau S, Carniel E, et al. Multidrug resistance in Yersinia pes- Bugs in Southern MDR plasmid from Salmonella spp. tis mediated by a transferable plasmid. N to Y. pestis seems possible. However, Engl J Med.1997;337:677–80. Patagonia, we emphasize that to date no evidence 5. Hinnebusch BJ, Rosso M-L, Schwan TG, Argentina supports this type of event. Carniel E. High-frequency conjugative transfer of antibiotic resistance genes to Yersinia pestis in the fl ea midgut. Mol Mi- To the Editor: Triatoma infestans crobiol. 2002;46:349–54. bugs, the main vector of Chagas dis- The Centers for Disease Control and 6. Welch TJ, Fricke WF, McDermott PF, ease, historically occupied a large area Prevention provided many of the DNA White DG, Rosso M-L, Rasko DA, et from northeastern Brazil to Chubut samples. al. Multiple antimicrobial resistance in plague: an emerging public health risk. Province in Patagonia, Argentina (1). This work was funded by National PLoS One. 2007;2:e309. DOI: 10.1371/ Large-scale insecticide spraying dur- Institutes of Health, National Institute of journal.pone.0000309 ing the 1980s and 1990s reduced its Allergy and Infectious Diseases (grant 7. Centers for Disease Control and Preven- geographic range and abundance and tion. Imported plague–New York City, nos. AI070183 and AI30071); the Pacifi c 2002. MMWR Morb Mortal Wkly Rep. interrupted transmission of Trypano- Southwest Regional Center of Excellence 2003;52:725–8. soma cruzi, mainly in Uruguay, Chile, (grant no. AI065359); the Department of 8. Carattoli A, Bertini A, Villa L, Falbo V, and Brazil (2). However, T. infestans Homeland Security Science and Technolo- Hopkins KL, Threlfall EJ. Identifi cation and transmission of T. cruzi persist in of plasmids by PCR-based replicon typ- gy Directorate (grant nos. NBCH2070001 ing. J Microbiol Methods. 2005;63:219– the Gran Chaco, a large ecoregion in and HSHQDC-08-C-00158); and the 28. DOI: 10.1016/j.mimet.2005.03.018 Argentina, Bolivia, and Paraguay (3). Cowden Endowment in Microbiology at 9. Eskey CR, Prince FM, Fuller FB. Double Chubut Province has historically Northern Arizona University. infection of the rat fl eas X. cheopis and been an area with no risk for vector- N. fasciatus with Pasteurella and Salmo- nella. Public Health Rep (1896–1970). mediated transmission of T. cruzi; David M. Wagner, 1951;66:1318. only its extreme northern region was Janelle Runberg, Amy J. Vogler, 10. Silbergeld EK, Graham J, Price LB. categorized as having a low transmis- Judy Lee, Elizabeth Driebe, Industrial food animal production, sion risk (4,5). However, increased Lance B. Price, antimicrobial resistance, and hu- man health. Annu Rev Public Health. immigration from disease-endemic David M. Engelthaler, 2008;29:151–69. DOI: 10.1146/annurev. rural areas in Argentina and Bolivia W. Florian Fricke, Jacques Ravel, publhealth.29.020907.090904 into Chubut has raised concerns about and Paul Keim accidental introduction of T. infestans Author affi liations: Northern Arizona Univer- Address for correspondence: David M. Wagner, in travelers’ luggage (1) and establish- sity, Flagstaff, Arizona, USA (D.M. Wagner, Center for Microbial Genetics and Genomics, ment of a transmission cycle. J. Runberg, A.J. Vogler, J. Lee, P. Keim); Northern Arizona University, Flagstaff, AZ In January 2007, we conducted Translational Genomics Research Institute, 86011-4073, USA; email: dave.wagner@nau. a province-wide survey of 21 vil- Flagstaff (E. Driebe, L.B. Price, D.M. En- edu lages in Chubut Province previously gelthaler, P. Keim); and University of Mary- infested with T. infestans bugs by us- land School of Medicine, Baltimore, Mary- ing 0.2% tetramethrin as a dislodgant land, USA (W.F. Fricke, J. Ravel) agent (1 person-hour/house); no T. DOI: 10.3201/eid1605.090892 infestans bugs were detected (online Appendix Figure, www.cdc.gov/EID/ References content/16/4/887-appF.htm). Only T. patagonica bugs were found in 11% 1. Perry RD, Fetherston JD. Yersinia pestis— of peridomestic structures, and none etiologic agent of plague. Clin Microbiol Rev. 1997;10:35–66. were infected with T. cruzi. In June 2. Galimand M, Carniel E, Courvalin P. Re- 2007, a T. infestans–like bug was sistance of Yersinia pestis to antimicrobial found in a primary healthcare center agents. Antimicrob Agents Chemother. in Comodoro Rivadavia (45°51′S, 2006;50:3233–6. 3. Guiyoule A, Gerbaud G, Buchrieser C, 67°28′W), a city in southern Chubut Galimand M, Rahalison L, Chanteau S, Province (online Appendix Figure). et al. Transferable plasmid–mediated Healthcare center staff reported visits resistance to streptomycin in a clini- by immigrants from Bolivia a few days cal isolate of Yersinia pestis. Emerg In- fect Dis. 2001;7:43–8. DOI: 10.3201/ before this fi nding and suspected them eid0701.010106 to be the source. The bug was identi-

Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 16, No. 5, May 2010 887 LETTERS

fi ed morphologically as a T. infestans x of the bug from southern Patagonia (Salta and Jujuy), western (Mendoza, female and it laid 6 eggs. PCR ampli- was found only in Argentina and was San Juan, and San Luis), and southern fi cation of kinetoplast DNA showed not closely related to haplotypes from Argentina (Rio Negro), i.e., from the 3 that it was not infected by T. cruzi. Bolivia. putative sources of the bug. These im- DNA sequence analysis is useful We investigated the geographic migrants typically pay extended visits for investigating evolutionary history origin of non-native putative attend- to their home towns at least once a year and population structure within Tri- ees of the healthcare center in San and transport luggage in which the bug atominae (6). T. infestans bugs from Cayetano. These persons were immi- could have traveled. In 2006, San Juan Bolivia and Argentina showed ge- grants from Bolivia and from northern had the highest levels of domestic and netic differences for nuclear (7) and mitochondrial markers (6), including mitochondrial cytochrome oxidase I (mtCOI) (8). We used our mtCOI hap- lotype database, which includes pub- lished (8) and new domestic, perido- mestic, and sylvatic T. infestans from 65 locations in 13 provinces in Argen- tina (n = 346) and 3 departments in Bo- livia (n = 144), to analyze the mtCOI sequence of the bug found in southern Patagonia and determine if it could be assigned to a known haplotype from Bolivia or Argentina. We investigated phylogenetic relationships with other haplotypes by using neighbor-joining and Bayesian approaches. Our mtCOI database included 53 haplotypes: 42 were found in Argen- tina, 9 in Bolivia, and 2 in both coun- tries (Figure). The bug from southern Patagonia had haplotype x, which has been found in only 3 western or south- ern provinces in Argentina (San Juan, San Luis, and Rio Negro) (8; online Appendix Figure). Results of phylogenetic analyses were congruent (Figure). The neigh- bor-joining tree showed that haplo- type x formed a cluster with haplotype Figure. Phylogenetic relationships between mitochondrial cytochrome oxidase I gene h (Argentina) and haplotypes from haplotypes of Triatoma infestans from Argentina and Bolivia. The neighbor-joining tree was Bolivia clustered in 3 other groups: constructed by using MEGA 4.1 (www.megasoftware.net) and bootstrap values (based on 1,000 replications) >50% are shown above the branches. A Bayesian maximum clade 1) two groups with bootstrap values credibility tree was similar, and clade posterior probabilities >50% are shown below the >70% (one with haplotypes at, n, c, branches of the neighbor-joining tree. MRBAYES 3.1 (http://mrbayes.csit.fsu.edu) default and 33 haplotypes from Argentina, priors were assumed and run for 4 million generations. Convergence of the Markov chain and the other with haplotypes ab, ac, Monte Carlo analysis was investigated with the SD of split frequencies and diagnostics ad, ae, ap, and az); and 2) one group implemented in AWTY (http://ceb.csit.fsu.edu/awty). The model of evolution (Hasegawa- Kishino-Yano + invariable sites+ Γ [HKY + I + Γ]) was chosen with Mrmodeltest 2.3 with a bootstrap value of 68% (hap- (www.abc.se/∼nylander). Because MEGA 4.1 does not support HKY; the more inclusive lotypes ax and aa). The Bayesian tree Tamura-Nei method (www.megasoftware.net/WebHelp/part_iv___evolutionary_analysis/ showed that haplotypes from Bolivia computing_evolutionary_distances/distance_models/nucleotide_substitution_models/ were arranged in 2 well-supported hc_tamura_nei_distance.htm) was used for the neighbor-joining analysis. Haplotypes al, clades (posterior probabilities >83%) an, ao, ap, aq, at, au, ax, az, aaa, and aab are reported. DNA sequences are available in GenBank (accession nos. EF451005-EF451041, FJ439768, FJ811845–8, and GQ and that haplotype x was not included 478993–GQ 479005). *Two provinces in Argentina; †Tarija, Bolivia; ‡10 provinces in within any of them. Thus, haplotype Argentina. Scale bar indicates nucleotide substitutions per site.

888 Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 16, No. 5, May 2010 LETTERS peridomestic infestation with T. in- Romina V. Piccinali, Delmi M. 8. Piccinali RV, Marcet PL, Noireau F, festans (35% and 21%, respectively), Canale, Alejandra E. Sandoval, Kitron U, Gürtler RE, Dotson EM. Mo- lecular population genetics and phylo- including urban infestation (9). Men- Marta V. Cardinal, Oscar geography of the Chagas disease vector doza (not in our database) had con- Jensen, Uriel Kitron, and Triatoma infestans in South America. J siderable domestic and peridomestic Ricardo E. Gürtler Med Entomol. 2009;46:796–809. DOI: 10.1603/033.046.0410 infestations (both 7%), and San Luis Author affi liations: Universidad de Buenos 9. Carrizo Páez R, Pickenhayn J, Carrizo (0.5% and 5.3%, respectively) and Rio Aires, Buenos Aires, Argentina (R.V. Pic- Páez M. Chagas urbano en San Juan. Di- Negro (both <0.1%) had low infesta- cinali, M.V. Cardinal, R.E. Gürtler); Coor- agnóstico, revisión y propuesta para un tions in 2001 (4) and thereafter (C. dinación Nacional de Control de Vectores, sistema integrado de ataque. Revista Ar- gentina Cardiologia. 2008;76:480–7. Spillmann, unpub. data). Bolivia, Sal- Córdoba, Argentina (D.M. Canale); Sec- 10. Gurevitz JM, Ceballos LA, Kitron U, ta, and Jujuy are excluded as potential retaría de Salud de Chubut, Chubut, Ar- Gürtler RE. Flight initiation of Triatoma sources of the bug because haplotypes gentina (A.E. Sandoval, O. Jensen); and infestans (Hemiptera: Reduviidae) under closely related to haplotype x were not Emory University, Atlanta, Georgia, USA natural climatic conditions. J Med Ento- mol. 2006;43:143–50. DOI: 10.1603/0022- found in these places. Active dispersal (U. Kitron) 2585(2006)043[0143:FIOTIH]2.0.CO;2 from a local source can be ruled out DOI: 10.3201/eid1605.091260 because there is no precedent for T. Address for correspondence: Romina V. infestans in Comodoro Rivadavia, and References Piccinali, Laboratorio de Eco-Epidemiología, the mean temperature in June (8°C) is Departamento de Ecología, Genética y below the known threshold for fl ight 1. Lent H, Wygodzinsky P. Revision of the Evolución, Facultad de Ciencias Exactas Triatominae (Hemiptera: Reduviidae) and initiation (23°C) (10). y Naturales, Universidad de Buenos Aires, Our results show that molecular their signifi cance as vectors of Chagas’ disease. Bulletin of the American Museum Pabellón 2, Ciudad Universitaria, Int. Güiraldes phylogenetics can identify passive of Natural History. 1979;163:127–520. 2160, C1428EGA, Buenos Aires, Argentina; transport of insects into areas where 2. Schofi eld CJ, Jannin J, Salvatella R. The email: [email protected] a disease is not endemic and rule out future of Chagas disease control. Trends putative sources supported only by Parasitol. 2006;22:583–8. DOI: 10.1016/j. pt.2006.09.011 circumstantial evidence. Our fi ndings 3. Gürtler RE, Kitron U, Cecere MC, Segura reinforce the need for sustained and EL, Cohen JE. Sustainable vector con- coordinated vector surveillance and trol and management of Chagas disease control at a regional level (3). in the Gran Chaco, Argentina. Proc Natl Acad Sci U S A. 2007;104:16194–9. DOI: 10.1073/pnas.0700863104 Acknowledgments 4. Pan American Health Organization. XI Serologic Survey We thank Mirko Rojas-Cortez for Reunión de la Comisión Interguberna- of Hantavirus providing samples from Bolivia, Ricardo mental para la Eliminación de Triatoma infestans y la Interrupción de la Tripano- Infection, Brazilian Váez for assisting in bug searches, Fer- somiasis Americana por Transfusión, nando Garelli and Juan Gurevitz for care- Paraguay. Iniciativa de salud del cono Sur Amazon fully reading the manuscripts and sugges- (INCOSUR). 2002 March [cited 2009 Oct To the Editor: Since the etiol- tions, and Marcela Rodriguero for helpful 13]. http://www.paho.org/spanish/HCP/ HCT/DCH/xi-incosur.htm ogy of hantavirus cardiopulmonary discussions on an earlier version of the 5. Segura EL, Cura EN, Estani SA, An- syndrome (HCPS) was recognized in manuscript. drade J, Lansetti JC, de Rissio AM, et al. Long-term effects of a nationwide control 1993 in the United States (1), vari- This study was supported by research program on the seropositivity for Try- ous hantaviruses have been associated grant no. R01 TW05836 from the National panosoma cruzi infection in young men with the syndrome in South America Institutes of Health Fogarty International from Argentina. Am J Trop Med Hyg. (2,3). Depending on the viral geno- 2000;62:353–62. Center and the National Institute of En- 6. Abad-Franch F, Monteiro FA. Molecu- type involved, hantavirus infection vironmental Health Sciences to U.K. and lar research and the control of Chagas can take a wide variety of forms, from R.E.G., and by grants from the Universi- disease vectors. An Acad Bras Cienc. asymptomatic or oligosymptomatic to dad de Buenos Aires and Agencia Nacio- 2005;77:437–54. DOI: 10.1590/S0001- the classic clinical form (4,5). 37652005000300007 nal de Promoción Científi ca y Técnica 7. Bargues MD, Klisiowicz DR, Panzera F, The fi rst cases of HCPS in Bra- (Argentina) to R.E.G. R.V.P., M.V.C., and Noireau F, Marcilla A, Perez R, et al. Ori- zil were reported in the state of São R.E.G. are scientifi c investigators of Con- gin and phylogeography of the Chagas dis- Paulo in 1993 (6,7). In 2000, an out- sejo Nacional de Investigaciones Científi - ease main vector Triatoma infestans based break of HCPS was reported in the on nuclear rDNA sequences and genome cas y Técnicas (Argentina). size. Infect Genet Evol. 2006;6:46–62. municipality of Anajatuba in the state DOI: 10.1016/j.meegid.2005.01.006 of Maranhão in the Maranhão western

Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 16, No. 5, May 2010 889