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Screening and genotyping of Orientia tsutsugamushi from field-collected on-host chiggers (Acari: Prostigmata) recovered from a positive scrub typhus locality in Kelantan, Malaysia

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Abstract

Orientia tsutsugamushi is the causative agent of scrub typhus vectored by larval stages of trombiculid mites (chiggers) that occur in most tropical regions of Southeast Asia. A total of 242 chiggers extracted from eight small mammals captured from a positive scrub typhus locality in Kelantan, Malaysia, were screened for the presence of O. tsutsugamushi. The chiggers were grouped in 16 pools for extraction of DNA prior to screening of O. tsutsugamushi based on the nucleotide sequence of 56-kDa type specific antigen (TSA) gene using nested polymerase chain reaction. Two species of on-host chiggers were identified, the one, Leptotrombidium deliense, much more dominant (94.8%) than the other, Ascoshoengastia sp. (5.2%). The pathogen was detected in two pools (12.5%) of L. deliense recovered from Rattus rattus and Tupaia sp. The 56-kDa TSA gene sequence analysis revealed the O. tsutsugamushi harboured in those chiggers were Karp prototype strain with high similarity (99.3%). Findings of this study strongly supported the existence of scrub typhus infections in certain parts of Malaysia which agrees with previous local reports. Moreover, this study highlighted the pressing need of a large-scale close observation of O. tsutsugamushi DNA sequences from chiggers that can probably be collected from other positive scrub typhus localities to precisely provide the distribution and prevalence of this zoonotic pathogen.

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References

  • Bonell A, Lubell Y, Newton PN, Crump JA, Paris DH (2017) Estimating the burden of scrub typhus: a systematic review. PLoS Negl Trop Dis 11(9):e0005838

    Article  PubMed  PubMed Central  Google Scholar 

  • Brown GW, Robinson DM, Huxsoll DL, Ng TS, Lim KJ (1976) Scrub typhus: a common cause illness in indigenous populations. Trans R Soc Trop Med Hyg 70(5–6):444–448

    Article  CAS  PubMed  Google Scholar 

  • Chaisiri K, Cosson JF, Morand S (2017) Infection of rodents by Orientia tsutsugamushi, the agent of scrub typhus, in relation to land use in Thailand. Trop Med Infect Dis 2(4):53

    Article  PubMed Central  Google Scholar 

  • Chaisiri K, Christina-Gill A, Stekolnikov AA, Hinjoy S, McGarry JW, Darby AC, Morand S, Makepeace BL (2019) Ecological and microbiological diversity of chigger mites, including vectors of scrub typhus, on small mammals across stratified habitats in Thailand. Animal Microbiome 1:18

    Article  PubMed  PubMed Central  Google Scholar 

  • Chakraborty S, Sarma N (2017) Scrub typhus: an emerging threat. Indian J Dermatol 62(5):478–485

    PubMed  PubMed Central  Google Scholar 

  • Ching WM, Rowland D, Zhang Z, Bourgeois AL, Kelly D, Dasch GA, Devine PL (2001) Early diagnosis of scrub typhus with a rapid flow assay using recombinant major outer membrane protein antigen (r56) of Orientia tsutsugamushi. Clin Diagn Lab Immunol 8(2):409–414

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cosson JF, Galan M, Bard E, Razzauti M, Bernard M, Morand S, Brouat C, Dalecky A, Charbonnel N, Vayssier-Taussat M (2015) Detection of Orientia sp DNA in rodents from Asia West Africa and Europe. Parasit Vectors 8:172

    Article  PubMed  PubMed Central  Google Scholar 

  • Elliot I, Pearson I, Dahal P, Thomas NV, Roberts T, Newton PN (2019) Scrub typhus ecology: a systematic review of Orientia in vectors and hosts. Parasit Vectors 12:513

    Article  Google Scholar 

  • Fletcher W, Field JW (1927) The tsutsugamushi disease in the Federated Malay States. Bull Inst Med Res 1:1–26

    Google Scholar 

  • Fournier PE, Siritantikorn S, Rolain JM, Suputtamongkol Y, Hoontrakul S, Charoenwat S, Losuwanaluk K, Parola P, Raoult D (2008) Detection of new genotypes of Orientia tsutsugamushi infecting humans in Thailand. Clin Microb Infect 14(2):168–173

    Article  Google Scholar 

  • Frances SP, Watcharapichat P, Phulsuksombati D, Tanskul P (2001) Investigation of the role of Blankaartia acuscutellaris (Acari: Trombiculidae) as a vector of scrub typhus in central Thailand. Southeast Asian J Trop Med Public Health 32(4):863–866

    CAS  PubMed  Google Scholar 

  • Francis CM (2019) Field guide to the mammals of South-east Asia (2nd Edition). Bloomsbury Wildl, London, p 416

    Google Scholar 

  • Furuya Y, Yoshida Y, Katayama T, Yamamoto S, Kawamura A (1993) Serotype-specific amplification of Rickettsia tsutsugamushi DNA by nested polymerase chain reaction. J Clin Microbiol 31(6):1637–1640

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Window 95/98/NT. Nucleic Acids Symp Ser 41:95–98

    CAS  Google Scholar 

  • Hanifah AL, Mariana A, Vishalani VN, Ho TM (2013) Detection of Orientia tsutsugamushi in chiggers and tissue of small mammals using Polymerase Chain Reaction. The Exp 11(2):683–689

    Google Scholar 

  • Huxsoll DL (1978) Scrub typhus- A problem in Malaysia? Malaysian J Path 1:1–6

    Google Scholar 

  • Kala D, Gupta S, Nagraik R, Verma V, Thakur A, Kaushal A (2020) Diagnosis of scrub typhus: recent advancements and challenges. 3 Biotech 10(9):396

    Article  PubMed  PubMed Central  Google Scholar 

  • Kelly DJ, Fuerst PA, Ching WM, Richards AL (2009) Scrub typhus: the geographic distribution of phenotypic and genotypic variants of Orientia tsutsugamushi. Clin Infect Dis 48:S203–S230

    Article  CAS  PubMed  Google Scholar 

  • Kim DM, Yun NR, Yang TY, Lee JH, Yang JT, Shim SK, Choi EN, Park MY, Lee SH (2006) Usefulness of nested PCR for diagnosis of scrub typhus in clinical practice: a prospective study. Am J Trop Med Hyg 75(3):542–545

    Article  PubMed  Google Scholar 

  • Kocher C, Jiang J, Morrison AC, Castillo R, Leguia M, Loyola S, Ampuero S, Bausch DG, Richard AL (2017) Scrub typhus in the Peruvian Amazon Basin. Emerg Infect Dis 23:1389–1391

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Koh GC, Maude RJ, Paris DH, Newton PN, Blacksell SD (2010) Diagnosis of scrub typhus. Am J Trop Med Hyg 82:368–370

    Article  PubMed  PubMed Central  Google Scholar 

  • Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33(7):1870–1874

    Article  CAS  Google Scholar 

  • Kuo CC, Lee PL, Chen CH, Wang HC (2015) Surveillance of potential hosts and vectors of scrub typhus in Taiwan. Parasit Vectors 8:611

    Article  PubMed  PubMed Central  Google Scholar 

  • Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23:2947–2948

    Article  CAS  Google Scholar 

  • Lerdthusnee K, Khlaimanee N, Monkanna T, Sangjun N, Mungviriya S, Linthicum KJ, Frances SP, Kollars TM Jr, Coleman RE (2002) Efficiency of Leptotrombidium chiggers (Acari: Trombiculidae) at transmitting Orientia tsutsugamushi to laboratory mice. J Med Entomol 39(3):521–525

    Article  PubMed  Google Scholar 

  • Lin PR, Tsai HP, Tsui PY, Weng MH, Kuo MD, Lin HC, Chen KC, Ji DD, Chu DM, Liu WT (2011) Genetic typing, based on the 56-Kilodalton type-specific antigen gene, of Orientia tsutsugamushi strains isolated from chiggers collected from wild-caught rodents in Taiwan. Appl Envt Microbiol 77(10):3398–3405

    Article  CAS  Google Scholar 

  • Lv Y, Guo XG, Jin DC (2018) Research progress on Leptotrombidium deliense. Korean J Parasitol 56(4):313–324

    Article  PubMed  PubMed Central  Google Scholar 

  • Lyu Y, Tian L, Zhang L, Dou X, Wang X, Li W, Zhang X, Sun Y, Guan Z, Li X, Wang Q (2013) A case-control study of risk factors associated with scrub typhus infection in Beijing China. PLoS ONE 8(5):e63668

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mariana A, Mohd Kulaimi B, Suhaili ZA, Shahrul-Anuar MS, Zalipah MN, Ho TM (2011) Acarine ectoparasites of Panti Forest Reserve in Johore Malaysia. Asian Pac J Trop Biomed 1(1):1–5

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Medway L (1983) The wild mammals of Malaya (Peninsular Malaysia and Singapore). Oxford University Press, Kuala Lumpur

    Google Scholar 

  • Nadchatram M (1970) Correlation of habitat, environment and color of chiggers, and their potential significance in the epidemiology of scrub typhus in Malaya (Prostigmata: Trombiculidae). J Med Entomol 7:131–144

    Article  CAS  PubMed  Google Scholar 

  • Nadchatram M (2008) The beneficial rain forest ecosystem with environmental effects on zoonoses involving ticks and mites, a Malaysian perspective and review. Trop Biomed 25(2):1–92

    Google Scholar 

  • Nadchatram M, Dohany AL (1974) A pictorial key to the subfamilies, genera and subgenera of Southeast Asian chiggers. Bulletin Inst Med Res Malays 16:1–67

    Google Scholar 

  • Osuga K, Kimura M, Goto H, Shimada K, Suto T (1991) A case of tsutsugamushi disease probably contracted in Africa. Eur J Clin Microbiol Infect Dis 10:95–96

    Article  CAS  PubMed  Google Scholar 

  • Paramasvaran S, Sani RA, Hassan L, Krishnasamy M, Jeffery J, Oothuman P, Salleh I, Lim KH, Sumarni MG, Santhana RL (2009) Ectoparasite fauna of rodents and shrews from four habitats in Kuala Lumpur and the states of Selangor and Negeri Sembilan, Malaysia and its public health significance. Trop Biomed 26(3):303–311

    CAS  PubMed  Google Scholar 

  • Paris DH, Shelite TR, Day NP, Walker D (2013) Unresolved problem related to scrub typhus: a seriously neglected life-threatening diseases. Am J Trop Med Hyg 89:301–307

    Article  PubMed  PubMed Central  Google Scholar 

  • Patricia KA, Hoti SL, Kanungo R, Jambulingam P, Shashikala N, Naik AC (2017) Improving the diagnosis of scrub typhus by combining groEL based polymerase chain reaction and IgM ELISA. J Clin Diag Res 11(8):27–313

    CAS  Google Scholar 

  • Payne J, Francis CM, Phillips K (2005) A field guide to the mammals of Borneo. Sabah Society, Kota Kinabalu, p 332

    Google Scholar 

  • Phommasone K, Paris DH, Anantatat T, Castonguay-Vanier J, Keomany S, Souvannasing P, Blacksell SD, Mayxay M, Newton PN (2013) Concurrent infection with murine typhus and scrub typhus in Southern Laos—the mixed and the unmixed. PLoS Negl Trop Dis 7(8):e2163

    Article  PubMed  PubMed Central  Google Scholar 

  • Rajapakse S, Weeratunga P, Sivayoganathan S, Fernando SD (2017) Clinical manifestation of scrub typhus. Trans R Soc Trop Med Hyg 111:43–54

    Article  PubMed  Google Scholar 

  • Richard AL, Jiang J (2020) Scrub typhus: historic perspective and status of the worldwide presence of Orientia species. Trop Med Infect Dis 5(2):49

    Article  Google Scholar 

  • Rodkvamtook W, Ruang-areerate T, Gaywee J, Richards AL, Jeamwattanalert P, Bodhidatta D, Sangjun N, Prasartvit A, Jatisatienr A, Jatisatienr C (2011) Isolation and characterization of Orientia tsutsugamushi from rodents captured following a scrub typhus outbreak at a Military Training Base, Bothong District, Chinburi Province, Central Thailand. Am J Trop Med Hyg 84(4):599–607

    Article  PubMed  PubMed Central  Google Scholar 

  • Ruang-areerate T, Jeamwattanalert P, Rodkvamtook W, Richards AL, Sunyakumthorn P, Gaywee J (2011) Genotype diversity and distribution of Orientia tsutsugamushi causing scrub typhus in Thailand. J Clin Microbiol 49(7):2584–2589

    Article  PubMed  PubMed Central  Google Scholar 

  • Santibañez P, Palomar A, Portillo A, Santibañez S, Oteo JA (2015) The role of chiggers as human pathogens. In: Samie A (ed) An Overview of Tropical Diseases. Intech Open Limited, London, pp 173–202

    Google Scholar 

  • Saraswati K, Day NP, Mukaka M, Blacksell SD (2018) Scrub typhus point-of-care testing: a systematic review and meta-analysis. PLoS Negl Trop Dis 12:e0006330

    Article  PubMed  PubMed Central  Google Scholar 

  • Sasa M (1954) Comparative epidemiology of Tsutsugamushi disease in Japan. Jpn J Exp Med 24:335–361

    CAS  PubMed  Google Scholar 

  • Shirai A, Dohany AL, Ram S, Chiang GL, Huxsoll DL (1981) Serological classification of Rickettsia tsutsugamushi organisms found in chiggers (Acarina: Trombiculidae) collected in Peninsular Malaysia. Trans R Soc Trop Med Hyg 75:580–582

    Article  CAS  PubMed  Google Scholar 

  • Shirai A, Robinson DM, Brown GW, Gan E, Huxsoll DL (1979) Antigenic analysis by direct immuno-fluorecence of 114 isolates of Rickettsia tsutsugamushi recovered from febrile patients in rural Malaysia. Jpn J Med Sci Biol 32:337–344

    Article  CAS  PubMed  Google Scholar 

  • Shivalli S (2016) Diagnostic evaluation of rapid tests for scrub typhus in the Indian population is needed. Infect Dis Poverty 5:40

    Article  PubMed  PubMed Central  Google Scholar 

  • Takhampunya R, Korkusol A, Promsathaporn S, Tippayachai B, Leepitakrat S, Richards AL, Davidson SA (2018) Heterogeneity of Orientia tsutsugamushi genotypes in field-collected trombiculid mites from wild-caught small mammals in Thailand. PLoS Negl Trop Dis 12(7):e0006632

    Article  PubMed  PubMed Central  Google Scholar 

  • Takhampunya R, Tippayachai B, Promsathaporn S, Leepitakrat S, Monkanna T, Schuster AL, Melendrez MC, Paris DH, Richards AL, Richardson JH (2014) Characterization based on the 56-kDa type-specific antigen gene of Orientia tsutsugamushi genotypes isolated from Leptotrombidium mites and the rodents host post-infection. Am J Trop Med Hyg 90(1):139–146

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tanskul P, Strickman D, Eamsila C, Kelly DJ (1994) Rickettsia tsutsugamushi in chiggers (Acari: Trombiculidae) associated with rodents in central Thailand. J Med Entomol 31(2):225–230

    Article  CAS  PubMed  Google Scholar 

  • Tantibhedhyangkul W, Wongsawat E, Silpasakorn S, Waywa D, Saenyasiri N, Suesuay J, Thipmontree W, Suputtamongkol Y (2017) Use of multiplex real-time PCR to diagnose scrub typhus. J Clin Microbiol 55(5):1377–1387

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tay ST, Mohamed Zan HA, Lim YAL, Ngui R (2013) Antibody prevalence and factors associated with exposure to Orientia tsutsugamushi in different aboriginal subgroups in West Malaysia. PLoS Negl Trop Dis 7(8):e2341

    Article  PubMed  PubMed Central  Google Scholar 

  • Tay ST, Ho TM, Rohani MY, Devi S (2000) Antibodies to Orientia tsutsugamushi, Rickettsia typhi and spotted fever group rickettsiae among febrile patients in rural areas of Malaysia. Trans R Soc Trop Med Hyg 94:280–284

    Article  CAS  PubMed  Google Scholar 

  • Taylor AJ, Paris DH, Newton PN (2015) A systematic review of mortality from untreated scrub typhus (Orientia tsutsugamushi). PLoS Negl Trop Dis 9(8):e0003971

    Article  PubMed  PubMed Central  Google Scholar 

  • TraubWisseman RCL Jr (1974) The ecology of chigger-borne rickettsioses (scrub typhus). J Med Entomol 11:237–303

    Article  Google Scholar 

  • Trung NV, Hi LT, Cuong DD, Ha DT, Hoa TM, Lien VN, Hoa NT (2019) Analysis of the 56-kDa type specific antigen gene of Orientia tsutsugamushi from northern Vietnam. PLoS ONE 14(8):e0221588

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wangrangsimakul T, Elliott I, Nedsuwan S, Kumlert R, Hinjoy S, Chaisiri K, Day NPJ, Morand S (2020) The estimated burden of scrub typhus in Thailand from national surveillance data (2003–2018). PLoS Negl Trop Dis 14(4):e0008233

    Article  PubMed  PubMed Central  Google Scholar 

  • Weitzel T, Dittrich S, Lopez J, Phuklia W, Martinez-Valdebenito C, Velasquez K, Blacksell SD, Paris DH, Abarca K (2016) Endemic scrub typhus in South America. N Engl J Med 375(10):954–961

    Article  PubMed  Google Scholar 

  • Yathramullage S, Rajapakse J, Boyagoda S (2018) Rickettsiae reservoirs among small mammals (rats, mice and shrews) and their arthropod vectors in Sri Lanka. Ceylon J Sci 47(2):175–183

    Article  Google Scholar 

  • Zan HAM, Ponnampalavanar S, Omar SFS, Savithiri PD, Lim YAL, Tay ST (2016) Genetic variants of Orientia tsutsugamushi identified from scrub typhus cases in Malaysia. Trop Med 33(1):203–208

    Google Scholar 

  • Zhang M, Zhao ZT, Yang HL, Zhang AH, Xu XQ, Meng XP, Zhang HY, Wang XJ, Li Z, Ding SJ, Yang L, Zhang LY (2013) Molecular epidemiology of Orientia tsutsugamushi in chiggers and ticks from domestic rodents in Shadong, northern China. Parasit Vectors 6:312

    Article  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

The authors would like to thank the Director-General of Health Malaysia for his permission to publish this article. We wish to thank the entomologists from Kelantan State Health Department, Ms Sharifah Nadwah Md Yunus, Ms Nur Akmarina Mohd Noordin and Ms Suraya Sulaiman for their assistance in the field and the collection of the samples.

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Authors and Affiliations

  1. Infectious Diseases Research Centre, Institute for Medical Research (IMR), National Institutes of Health, Ministry of Health Malaysia, Acarology Unit, Jalan Setia Murni U13/52, Seksyen U13, Setia Alam, 40170, Shah Alam, Selangor, Malaysia

    F. C. L. Ernieenor, A. Fadillah, J. Canedy & A. Mariana

  2. Disease Control Division, Entomology and Pest Unit, Kelantan State Health Department, Ministry of Health Malaysia, Jalan Mahmood, 15200, Kota Bharu, Kelantan, Malaysia

    M. J. NorJaiza

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  1. F. C. L. Ernieenor
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Ernieenor, F.C.L., NorJaiza, M.J., Fadillah, A. et al. Screening and genotyping of Orientia tsutsugamushi from field-collected on-host chiggers (Acari: Prostigmata) recovered from a positive scrub typhus locality in Kelantan, Malaysia. Exp Appl Acarol 84, 171–182 (2021). https://doi.org/10.1007/s10493-021-00609-3

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