Russian Journal of Infection and ImmunityRussian Journal of Infection and ImmunityИнфекция и иммунитет2220-76192313-7398SPb RAACI129510.15789/2220-7619-MTO-1295ORIGINAL ARTICLESОРИГИНАЛЬНЫЕ СТАТЬИMolecular typing of Rickettsia akariГенетическое типирование Rickettsia akariEremeevaM. E.ЕремееваМ. Е.
United States

Eremeeva Marina E., PhD, MD (Bioсhemistry), Professor, Laboratory Director, Jiann-Ping Hsu College of Public Health

PO Box 8015, Statesboro, GA 30458

Еремеева Марина Евгеньевна, доктор биологических наук, профессор, заведующий лабораторией, Колледж народного здравоохранения им. Цзянн-Пин Хсу

30458, Джорджия, Стейтсборо, почтовый ящик 8015

meremeeva@georgiasouthern.eduSturgeonM. M.СтуржеонМ. М.
United States

MPH, Training Fellow

Atlanta, Georgia

Магистр общественного здравоохранения, стажер

Атланта, Джорджия

mmsturgeon.mph@gmail.comWillardJ. K.УиллардД. К.
United States

BS, Training Fellow

Atlanta, Georgia

Бакалавр биологических наук, стажер

Атланта, Джорджия

jwilard004@gmail.comKarpathyS. E.КарпатиС. Э.
United States

PhD, Microbiologist

Atlanta, Georgia

Доктор биологических наук, микробиолог

Атланта, Джорджия

evu2@cdc.govMadanA.МаданА.
United States

PhD, Senior Research Scientist

Redmond, WA

Доктор биологических наук, старший научный сотрудник

Редмонд, Вашингтон

anupmadan@gmail.comDaschG. A.ДашГ. А.
United States

PhD, Leading Microbiologist and Group Leader

Atlanta, Georgia

Доктор биологических наук, ведущий микробиолог и руководитель группы

Атланта, Джорджия

ged4@cdc.govGeorgia Southern UniversityУниверситет Южной ДжорджииRickettsial Zoonoses Branch, Division of Vector-Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention (CDC)Центр Контроля и Профилактики ЗаболеванийCovance Genomics LaboratoryКованс Геномик лаборатория070820201034975050111201914032020Copyright ©; 2020, Eremeeva M.E., Sturgeon M.M., Willard J.K., Karpathy S.E., Madan A., Dasch G.A.Copyright ©; 2020, Еремеева М.Е., Стуржеон М.М., Уиллард Д.К., Карпати С.Э., Мадан А., Даш Г.А.2020Eremeeva M.E., Sturgeon M.M., Willard J.K., Karpathy S.E., Madan A., Dasch G.A.Еремеева М.Е., Стуржеон М.М., Уиллард Д.К., Карпати С.Э., Мадан А., Даш Г.А.https://creativecommons.org/licenses/by/4.0https://iimmun.ru/iimm/article/view/1295

Rickettsia akari, an obligately intracellular bacterium, is the causative agent of the cosmopolitan urban disease rickettsialpox. R. akari is an atypical representative of spotted fever group rickettsiae (SFG) as it is associated with rodent mites rather than ticks or fleas; however, only limited information is available about the degree of genetic variability found among isolates of R. akari. We examined 13 isolates of R. akari from humans, rodents and mites in the USA, the former Soviet Union, and the former Yugoslavia made between 1946 and 2003 for diversity in their tandem repeat regions (TR) and intergenic regions (IGR). The 1.23 Mb genome of R. akari strain Hartford CWPP was analyzed using Tandem Repeat Finder software (http://tandem.bu.edu) and 374 different TRs were identified, with size variation from 1 to 483 bp and with TR copy numbers ranging between 21 and 1.9, respectively. No size polymorphisms were detected among the 11 TR regions examined from 5 open reading frames and 6 IGR. Eighteen non-TR IGR’s were amplified and sequenced for the same isolates comprising a total of 5.995 bp (0.49%) of the Hartford CWPP strain chromosome. Three single nucleotide polymorphism (SNP) sites were detected in two IGR’s which permitted separation of the five R. akari isolates from Ukraine SSR from the other eight isolates. In conclusion, this is the first study reporting genetic heterogeneity among R. akari isolates of different geographic origins. Further exploration of this genetic diversity is needed to understand better the geographic distribution of R. akari and the epidemiology of rickettsialpox. The potential of mites as hosts for other rickettsial agents also needs further investigation.

Rickettsia akari является облигатной внутриклеточной бактерией и вызывает осповидный риккетсиоз. Rickettsia akari является атипичным представителем группы риккетсиозных пятнистых лихорадок (SFG), поскольку она циркулирует в гамазовых клещах; однако в отношении степени генетического разнообразия изолятов R. akari имеются лишь ограниченные данные. Мы изучили 13 изолятов R. akari, выделенных от человека, грызунов и клещей в США, странах бывшего СССР и бывшей Югославии за период с 1946 по 2003 г. на предмет разнообразия в составе тандемных повторов (TR) и межгенных районов (IGR). При помощи программы Tandem Repeat Finder software (http://tandem.bu.edu) по поиску тандемных повторов Tandem Repeat Finder (http://tandem.bu.edu) был проанализирован геном длиною 1,23 Мб штамма R. akari Hartford CWPP; обнаружены 374 различных TR размером от 1 до 483 п.н. и количестве копий TR колеблющихся от 21 до 1,9 соответственно. Среди 11 TR из 5 открытых рамок считывания и 6 IGR не обнаружено размерного полиморфизма. Восемнадцать не-TR IGR были амплифицированы и секвенированы из всех изолятов, составляющих в целом 5995 п.н. (0,49%) хромосомы штамма Hartford CWPP. Три точечные мутации (SNP) были обнаружены в двух IGR, что позволило отделить пять изолятов R. akari из Украинской ССР от восьми других изолятов. В заключение следует отметить, что настоящее исследование является первым описанием генетического разнообразия среди изолятов R. akari из различных географических регионов. Дальнейшее изучение генетического разнообразия необходимо для более точного понимания географического распространения R. akari и эпидемиологии осповидного риккетсиоза. Оценка потенциала гамазовых клещей в качестве хозяев для возбудителей других риккетсиозов также требует дополнительных исследований.

rickettsialpoxmiteLiponyssoides sanguineusRickettsia akaririckettsiaespotted fever group rickettsiaetandem repeatsintergenic regionsgenotypingmultilocus sequence typingосповидный риккетсиозгамазовые клещиLiponyssoides sanguineusRickettsia akariриккетсиигенотипированиетандемные повторымежгенные фрагментымультилокуснoe генотипированиеThis study was supported in part by the NIH grant AI05326-03. This research was supported in part by an appointment of M. Sturgeon to the Emerging Infectious Diseases (EID) Fellowship Program administered by the Association of Public Health Laboratories (APHL) and funded by the Centers for Disease Control and Prevention (CDC). The findings and conclusions in this article are those of the authors and do not necessarily represent the views of the Centers for Disease Control and Prevention. We thank the many investigators who contributed stocks of the isolates used in this investigation1. Киселев Р.И., Волчанецкая Г.И. Сплошная дератизация и дезинсекция как метод борьбы с заболеваемостью оспоподобным риккетсиозом // Журнал микробиологии, эпидемиологии и иммунобиологии. 1954. № 12. C. 28–33.2. Киселев Р.И., Жданов В.М., Александрова Н.Н. К клинико-эпидемиологической характеристике оспоподобного риккетсиоза // Сб. тр. Харьковского НИИ вакцин и сывороток имени Мечникова. 1954. Т. ХХ. C. 253–257.3. Кулагин С.М. К характеристике эндемических риккетсиозов // Журнал микробиологии, эпидемиологии и иммунобиологии. 1952. Т. 12. С. 3–10.4. Benson G. Tandem repeats finder: a program to analyze DNA sequences. Nucleic Acids Res., 1999, vol. 27, no. 2, pp. 573–580. doi: 10.1093/nar/27.2.5735. Brouqui P., Parola P., Fournier P.E., Raoult D. Spotted fever rickettsioses in southern and eastern Europe. FEMS Immunol. Med. Microbiol., 2007, vol. 49, no. 1, pp. 2–12. doi: 10.1111/j.1574-695X.2006.00138.x6. Brown L.D., Macaluso K.R. Rickettsia felis, an emerging flea-borne rickettsiosis. Curr. Trop. Med. Rep., 2016, vol. 3, pp. 27–39. doi: 10.1007/s40475-016-0070-67. Cheng X., Jin Y., Lao S., Huang C., Huang F., Jia P., Zhang L. Multispacer typing (MST) of spotted fever group rickettsiae isolated from humans and rats in Chengmai County, Hainan Province, China. Trop. Med. Int. Health., 2014, vol. 42, no. 3, pp. 107–114. doi: 10.2149/tmh.2014-038. Choi Y.J., Lee E.M., Park J.M., Lee K.M., Han S.H., Kim J.K., Lee S.H., Song H.J., Choi M.S., Kim I.S., Park K.H., Jang W.J. Molecular detection of various rickettsiae in mites (Acari: Trombiculidae) in southern Jeolla Province, Korea. Microbiol. Immunol., 2007, vol. 51, no. 3, pp. 307–312. doi: 10.1111/j.1348-0421.2007.tb03912.x9. Choi Y.J., Lee S.H., Park K.H., Koh Y.S., Lee K.H., Baik H.S., Choi M.S., Kim I.S., Jang W.J. Evaluation of PCR-based assay for diagnosis of spotted fever group rickettsiosis in human serum samples. Clin. Diagn. Lab. Immunol., 2005, vol. 12, no. 6, pp. 759–763. doi: 10.1128/CDLI.12.6.759-763.200510. Denison A.M., Amin B.D., Nicholson W.L., Paddock C.D. Detection of Rickettsia rickettsii, Rickettsia parkeri, and Rickettsia akari in skin biopsy specimens using a multiplex real-time polymerase chain reaction assay. Clin. Infect. Dis., 2014, vol. 59, no. 5, pp. 635–642. doi: 10.1093/cid/ciu35811. Eremeeva M., Balayeva N., Ignatovich V., Raoult D. Genomic study of Rickettsia akari by pulsed-field gel electrophoresis. J. Clin. Microbiol., 1995, vol. 33, no. 11, pp. 3022–3024.12. Eremeeva M.E., Balayeva N.M., Ignatovich V.F., Raoult D. Proteinic and genomic identification of spotted fever group rickettsiae isolated in the former USSR. J. Clin. Microbiol., 1993, vol. 31, no. 10, pp. 2625–2633.13. Eremeeva M.E., Bosserman E., Zambrano M., Demma L., Dasch G.A. Molecular typing of novel Rickettsia rickettsii isolates from Arizona. Ann. NY Acad. Sci., 2006, vol. 1078, pp. 573–577. doi: 10.1196/annals.1374.11414. Eremeeva M.E., Dasch G.A. Closing the gaps between genotype and phenotype in Rickettsia rickettsii. Ann. NY Acad. Sci., 2009, vol. 1166, pp. 12–26. doi: 10.1111/j.1749-6632.2009.04526.x15. Eustis E.B., Fuller H.S. Rickettsialpox. II. Recovery of Rickettsia akari from mites Allodermanyssus sanguineus, from West Hartford, Conn. Proceedings of the Society for Experimental Biology and Medicine Society for Experimental Biology and Medicine, 1952, vol. 80, no. 3, pp. 546–549. doi: 10.3181/00379727-80-1968516. Fournier P.E., Raoult D. Identification of rickettsial isolates at the species level using multi-spacer typing. BMC Microbiol., 2007, vol. 7, no. 7. doi: 10.1186/1471-2180-7-7217. Fournier P.E., Zhu Y., Ogata H., Raoult D. Use of highly variable intergenic spacer sequences for multispacer typing of Rickettsia conorii strains. J. Clin. Microbiol., 2004, vol. 42, no. 12, pp. 5757–5766. doi: 10.1128/JCM.42.12.5757-5766.200418. Frati F., Negri I., Fanciulli P.P., Pellecchia M., Dallai R. Ultrastructural and molecular identification of a new Rickettsia endosymbiont in the springtail Onychiurus sinensis (Hexapoda, Collembola). J. Invertebr. Pathol., 2006, vol. 93, no. 3, pp. 150–156. doi: 10.1016/j.jip.2006.07.00219. Gillespie J.J., Williams K., Shukla M., Snyder E.E., Nordberg E.K., Ceraul S.M., Dharmanolla C., Rainey D., Soneja J., Shallom J.M., Vishnubhat N.D., Wattam R., Purkayastha, A., Czar M., Crasta O., Setubal J.C., Azad A.F., Sobral B.S. Rickettsia phylogenomics: unwinding the intricacies of obligate intracellular life. PLoS One, 2008, vol. 3, no. 4: e2018. doi: 10.1371/journal.pone.000201820. Huang Y., Zhao L., Zhang Z., Liu M., Xue Z., Ma D., Sun X., Sun Y., Zhou C., Qin X., Zhu Y., Li W., Yu H., Yu X.J. Detection of a novel Rickettsia from Leptotrombidium scutellare mites (Acari: Trombiculidae) from Shandong of China. J. Med. Entomol., 2017, vol. 54, no. 3, pp. 544–549. doi: 10.1093/jme/tjw23421. Huebner R.J., Peggy S., Armstrong C. 1946b. Rickettsialpox: A newly recognized rickettsial disease: I. Isolation of the etiological agent. Public Health Rep., 1946, vol. 61, no. 45, pp. 1605–1614.22. Huebner R.J., Jellison W.L., Armstrong C. Rickettsialpox: A newly recognized rickettsial disease: V. Recovery of Rickettsia akari from a house mouse (Mus musculus). Public Health Rep., 1947, vol. 62, no. 22, pp. 777–780.23. Karpathy S.E., Dasch G.A., Eremeeva M.E. Molecular typing of isolates of Rickettsia rickettsii by use of DNA sequencing of variable intergenic regions. J. Clin. Microbiol., 2007, vol. 45, no. 8, pp. 2545–2553. doi: 10.1128/JCM.00367-0724. Kass E.M., Szaniawski W.K., Levy H., Leach J., Srinivasan K., Rives C. Rickettsialpox in a New York City hospital, 1980 to 1989. N. Engl. J. Med., 1994, vol. 331, no. 24, pp. 1612–1617. doi: 10.1056/NEJM19941215331240325. Krawczak F.S., Labruna M.B., Hecht J.A., Paddock C.D., Karpathy S.E. Genotypic characterization of Rickettsia bellii reveals distinct lineages in the United States and South America. Biomed Res. Int., 2018, 8505483. doi: 10.1155/2018/850548326. Krinsky W.L. Does epizootic lymphocytic choriomeningitis prime the pump for epidemic rickettsialpox? Rev. Infect. Dis., 1983, vol. 5, no. 6, pp. 1118–1119. doi: 10.1093/clinids/5.6.111827. Kumar S., Tamura K., Nei M. MEGA3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief. Bioinform., 2004, vol. 5, no. 2, pp. 150–163. doi: 10.1093/bib/5.2.15028. Maina A.N., Jiang J., Luce-Fedrow A., St John H.K., Farris C.M., Richards A.L. Worldwide presence and features of flea-borne Rickettsia asembonensis. Front. Vet. Sci., 2019, vol. 5, p. 334. doi: 10.3389/fvets.2018.0033429. Ozturk M.K., Gunes T., Kose M., Coker C., Radulovic S. Rickettsialpox in Turkey. Emerg. Infect. Dis., 2003, vol. 9, no. 11, pp. 1498–1499. doi: 10.3201/eid0911.03022430. Paddock C.D., Denison A.M., Lash R.R., Liu L., Bollweg B.C., Dahlgren F.S., Kanamura C.T., Angerami R.N., Pereira dos Santos F.C., Brasil Martines R., Karpathy S. Phylogeography of Rickettsia rickettsii genotypes associated with fatal Rocky Mountain spotted fever. Am. J. Trop. Med. Hyg., 2014, vol. 91, no. 3, pp. 589–597. doi: 10.4269/ajtmh.14-014631. Paddock C.D., Eremeeva M.E. Rickettsialpox. In: Rickettsial Diseases. Eds. Parola P., Raoult D. CRC Press, 2007, pp. 63–86.32. Paddock C.D., Koss T., Eremeeva M.E., Dasch G.A., Zaki S.R., Sumner J.W. Isolation of Rickettsia akari from eschars of patients with rickettsialpox. Am. J. Trop. Med. Hyg., 2006, vol. 75, no. 4, pp. 732–738.33. Paddock C.D., Zaki S.R., Koss T., Singleton J., Jr., Sumner J.W., Comer J.A., Eremeeva M.E., Dasch G.A., Cherry B., Childs J.E. Rickettsialpox in New York City: a persistent urban zoonosis. Annals of New York Academy of Sciences, 2003, vol. 990, pp. 36–44. doi: 10.1111/j.1749-6632.2003.tb07334.x34. Quevedo-Diaz M.A., Song C., Xiong Y., Chen H., Wahl L.M., Radulovic S., Medvedev A.E. Involvement of TLR2 and TLR4 in cell responses to Rickettsia akari. J. Leukoc. Biol., 2010, vol. 88, no. 4, pp. 675–685. doi: 10.1189/jlb.100967435. Radulovic S., Feng H.M., Morovic M., Djelalija B., Popov V., Crocquet-Valdes P., Walker D.H. Isolation of Rickettsia akari from a patient in a region where Mediterranean spotted fever is endemic. Clin. Infect. Dis., 1996, vol. 22, no. 2, pp. 216–220. doi: 10.1093/clinids/22.2.21636. Reeves W.K., Dowling A.P., Dasch G.A. Rickettsial agents from parasitic dermanyssoidea (Acari: Mesostigmata). Exp. Appl. Acarol., 2006, vol. 38, no. 2–3, pp. 181–188. doi: 10.1007/s10493-006-0007-137. Reeves W.K., Loftis A.D., Szumlas D.E., Abbassy M.M., Helmy I.M., Hanafi H.A., Dasch G.A. Rickettsial pathogens in the tropical rat mite Ornithonyssus bacoti (Acari: Macronyssidae) from Egyptian rats (Rattus spp.). Exp. Appl. Acarol., 2007, vol. 41, no. 1–2, pp. 101–107. doi: 10.1007/s10493-006-9040-338. Renvoise A., van’t Wout J.W., van der Schroeff J.G., Beersma M.F., Raoult D. A case of rickettsialpox in Northern Europe. Int. J. Infect. Dis., 2012, vol. 16, no. 3, pp. E221–E222. doi: 10.1016/j.ijid.2011.11.00939. Shapiro M.R., Fritz C.L., Tait K., Paddock C.D., Nicholson W.L., Abramowicz K.F., Karpathy S.E., Dasch G.A., Sumner J.W., Adem P.V., Scott J.J., Padgett K.A., Zaki S.R., Eremeeva M.E. Rickettsia 364D: a newly recognized cause of eschar-associated illness in California. Clin. Infect. Dis., 2010, vol. 50, no. 4, pp. 541–548. doi: 10.1086/64992640. Socolovschi C., Audoly G., Raoult D. Connection of toxin-antitoxin modules to inoculation eschar and arthropod vertical transmission in Rickettsiales. Comp. Immunol. Microbiol. Infect. Dis., 2013, vol. 36, no. 2, pp. 199–209. doi: 10.1016/j.cimid.2013.01.001.41. Vitorino L., de Sousa R., Bacellar F., Ze-Ze L. Characterization of a tandem repeat polymorphism in Rickettsia strains. J. Med. Microbiol., 2005, vol. 54, pt. 9, pp. 833–841. doi: 10.1099/jmm.0.45956-042. Vitorino L., de Sousa R., Bacellar F., Ze-Ze L. Automated method based in VNTR analysis for Rickettsiae genotyping. Ann. NY Acad. Sci., 2006, vol. 1078, pp. 582–586. doi: 10.1196/annals.1374.11643. Walker D.H., Hudnall S.D., Szaniawski W.K., Feng H.M. Monoclonal antibody-based immunohistochemical diagnosis of rickettsialpox: the macrophage is the principal target. Mod. Pathol., 1999, vol. 12, no. 5, pp. 529–533.44. Watson J. New building, old parasite: Mesostigmatid mites — an ever-present threat to barrier facilities. ILAR J., 2008, vol. 49, no. 3, pp. 303–309. doi: 10.1093/ilar.49.3.30345. Wikswo M.E., Hu R., Metzger M.E., Eremeeva M.E. Detection of Rickettsia rickettsii and Bartonella henselae in Rhipicephalus sanguineus ticks from California. J. Med. Entomol., 2007, vol. 44, no. 1, pp. 158–162. doi: 10.1603/0022-2585(2007)44[158:dorrab2.0.co;246. Zavala-Castro J.E., Zavala-Velázquez J.E., Peniche-Lara G.F., Sulú Uicab J.E. Human Rickettsialpox, Southeastern Mexico. Emerg. Infect. Dis., 2009, vol. 15, no. 10, pp. 1665–1667. doi: 10.3201/eid1510.08150747. Zhang L., Jin J., Fu X., Raoult D., Fournier P.E. 2006. Genetic differentiation of Chinese isolates of Rickettsia sibirica by partial ompA gene sequencing and multispacer typing. J. Clin. Microbiol., 2006, vol. 44, no. 7, pp. 2465–2467. doi: 10.1128/JCM.02272-0548. Zhu D.T., Xia W.Q., Rao Q., Liu S.S., Ghanim M., Wang X.W. Sequencing and comparison of the Rickettsia genomes from the whitefly Bemisia tabaci Middle East Asia Minor I. Insect Science, 2016, vol. 23, no. 4, pp. 531–542. doi: 10.1111/1744-7917.1236749. Zhu Y., Fournier P.E., Ogata H., Raoult D. Multispacer typing of Rickettsia prowazekii enabling epidemiological studies of epidemic typhus. J. Clin. Microbiol., 2005, vol. 43, no. 9, pp. 4708–4712. doi: 10.1128/JCM.43.9.4708-4712.2005