- ABSTRACT.
- INTRODUCTION
- MATERIALS AND METHODS
- RESULTS
- Clinical and histopathological data of atypical filariasis cases.
- Molecular characterization and phylogenetic analysis of filarial mt 12 S rRNA gene.
- Molecular characterization and phylogenetic analysis of filarial COI.
- Molecular characterization and phylogenetic analysis of filarial ITS1.
- Molecular detection and phylogenetic analysis of Wolbachia endosymbionts.
- DISCUSSION
- Supplemental Materials
- ACKNOWLEDGMENT
- REFERENCES
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
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Simón F , González-Miguel J , Diosdado A , Gómez PJ , Morchón R , Kartashev V , 2017. The complexity of zoonotic filariasis episystem and its consequences: A multidisciplinary view. BioMed Res Int 2017: 6436130.
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Somsap Y , Boonroumkaew P , Somsap A , Rodpai R , Sadaow L , Sanpool O , Maleewong W , Intapan P , 2021. Ocular dirofilariasis case in Thailand confirmed by molecular analysis to be caused by Dirofilaria immitis. Am J Trop Med Hyg 106: 204.
-
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Sukudom P , Phumee A , Siriyasatien P , 2018. First report on subconjunctival dirofilariasis in Thailand caused by a Dirofilaria sp. closely related to D. hongkongenesis. Acad J Sci Res 6: 114–116.
-
11.
Nunthanid P , Roongruanchai K , Wongkamchai S , Sarasombath PT , 2020. Case report: Periorbital filariasis caused by Brugia malayi. Am J Trop Med Hyg 103: 2336–2338.
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Thongpiya J , Sa-nguanraksa D , Samarnthai N , Sarasombath PT , 2020. Filariasis of the breast caused by Brugia pahangi: A concomitant finding with invasive ductal carcinoma. Parasitol Int 80: 102203.
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13.
Thongpiya J , Kreetitamrong S , Thongsit T , Toothong T , Rojanapanus S , Sarasombath P , 2021. The first case report of subcutaneous dirofilariasis caused by Dirofilaria repens in Thailand. Trop Parasitol 11: 125–127.
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Suphap N , Somkijrungroj T , Kongwattananon W , Supawatjariyakul W , Pataradool T , Kraivichian K , Jantarabenjakul W , Tulvatana W , Preativatanyou K , 2024. Ocular Brugia pahangi filariasis complicated by severe macular damage in Thailand: Case report and literature review. Am J Trop Med Hyg 110: 1158–1164.
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Wongkamchai S , Monkong N , Mahannol P , Taweethavonsawat P , Loymak S , Foongladda S , 2013. Rapid detection and identification of Brugia Malayi, B. Pahangi, and Dirofilaria immitis by high-resolution melting assay. Vector Borne Zoonotic Dis 13: 31–36.
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Casiraghi M , Favia G , Cancrini G , Bartoloni A , Bandi C , 2001. Molecular identification of Wolbachia from the filarial nematode Mansonella ozzardi. Parasitol Res 87: 417–420.
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Nuchprayoon S , Junpee A , Nithiuthai S , Chungpivat S , Suvannadabba S , Poovorawan Y , 2006. Detection of filarial parasites in domestic cats by PCR-RFLP of ITS1. Vet Parasitol 140: 366–372.
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Integrated Histological and Molecular Analysis of Filarial Species and Associated Wolbachia Endosymbionts in Human Filariasis Cases Presenting Atypically in Thailand
Patsharaporn T. Sarasombath
p.techasintana@gmail.com
Patsharaporn T. Sarasombath
Siriraj Integrative Center for Neglected Parasitic Diseases, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand;
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Panitta Sitthinamsuwan
Panitta Sitthinamsuwan
Department of Pathology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand;
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Sirirat Wijit
Sirirat Wijit
Siriraj Integrative Center for Neglected Parasitic Diseases, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand;
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Kedsara Panyasu
Kedsara Panyasu
Siriraj Integrative Center for Neglected Parasitic Diseases, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand;
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Kosol Roongruanchai
Kosol Roongruanchai
Siriraj Integrative Center for Neglected Parasitic Diseases, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand;
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Sukhum Silpa-Archa
Sukhum Silpa-Archa
Department of Ophthalmology, Rajavithi Hospital, College of Medicine, Rangsit University, Bangkok, Thailand;
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Matya Suwansirikul
Matya Suwansirikul
Buddhasothorn Hospital, Ministry of Public Health, Chachoengsao, Thailand;
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Peerasak Chortrakarnkij
Peerasak Chortrakarnkij
Division of Plastic and Reconstructive Surgery, Department of Surgery, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand;
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Pichet Ruenchit
Pichet Ruenchit
Siriraj Integrative Center for Neglected Parasitic Diseases, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand;
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Kanok Preativatanyou
Kanok Preativatanyou
Center of Excellence in Vector Biology and Vector-Borne Disease, Chulalongkorn University, Bangkok, Thailand;
Department of Parasitology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand
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Department of Parasitology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand
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Sirichit Wongkamchai
sirichit.won@mahidol.ac.th
Sirichit Wongkamchai
Siriraj Integrative Center for Neglected Parasitic Diseases, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand;
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ABSTRACT.
Atypical presentations of filariasis have posed diagnostic challenges due to the complexity of identifying the causative species and the difficulties in both diagnosis and treatment. In this study, we present the integrative histological and molecular analysis of seven atypical filariasis cases observed in regions of nonendemicity of Thailand. All filariasis cases were initially diagnosed based on histological findings. To confirm the causative species, molecular characterization based on both filarial mitochondrial (mt 12S rRNA and COI genes) and nuclear ITS1 markers was performed, together with the identification of associated Wolbachia bacterial endosymbionts. Among the cases studied, Brugia pahangi (N = 3), Brugia malayi (N = 1), Dirofilaria sp. “hongkongensis” (N = 2), and a suspected novel filarial species genetically related to Pelecitus copsychi (N = 1) were identified. By targeting the 16S rRNA gene, Wolbachia was also molecularly amplified in two cases of infection with Dirofilaria sp. “hongkongensis.” Phylogenetic analysis further revealed that the detected Wolbachia could be classified into supergroups C and F, indicating the high genetic diversity of this endosymbiont in Dirofilaria sp. “hongkongensis.” Furthermore, this study demonstrates the consistency between histological findings and species identification based on mitochondrial loci rather than on the nuclear ITS1. This suggests the utility of mitochondrial markers, particularly COI, as a highly sensitive and reliable diagnostic tool for the detection and differentiation of filarial species in clinical specimens. Precise identification of the causative species will facilitate accurate diagnosis and treatment and is also essential for the development of epidemiological and preventive strategies for filariasis.
- Supplemental Materials (PDF 86.462 KB)
Author Notes
Financial support: Siriraj Integrative Center for Neglected Parasitic Diseases (SiCNPD) is supported by a grant from the
Disclosures: The study protocol was approved by the Siriraj Institutional Review Board of Research involving human subjects (SIRB) (COA no. Si 397/2019). A waiver of consent was approved by the SIRB, as this research involved no more than minimal risk to the subjects. The waiver did not adversely affect the rights and welfare of the owners of the specimens.
Authors’ contributions: P. T. Sarasombath, K. Preativatanyou, P. Sitthinamsuwan, P. Ruenchit, and S. Wongkamchai conceived the idea of the study and designed the experiments. P. T. Sarasombath received grant funding. P. Sitthinamsuwan, S. Silpa-Archa, M. Suwansirikul, and P. Chortrakarnkij provided clinical data. P. T. Sarasombath, K. Preativatanyou, P. Sitthinamsuwan, S. Wijit, K. Panyasu, and K. Roongruanchai performed histological and molecular studies. P. T. Sarasombath and K. Preativatanyou performed sequence analysis and phylogenetic construction. P. T. Sarasombath and K. Preativatanyou analyzed the integrated data. P. T. Sarasombath, K. Preativatanyou, P. Ruenchit, and S. Wongkamchai wrote the first draft of the manuscript. P. T. Sarasombath and K. Preativatanyou revised the manuscript. All authors reviewed, finalized, and approved the final manuscript.
Current contact information: Patsharaporn T. Sarasombath, Siriraj Integrative Center for Neglected Parasitic Diseases, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand, E-mails: p.techasintana@gmail.com or patsharaporn.tec@mahidol.ac.th. Panitta Sitthinamsuwan, Department of Pathology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand, E-mails: panitta.sit@mahidol.ac.th or panitta.sit@gmail.com. Sirirat Wijit, Kedsara Panyasu, Kosol Roongruanchai, and Pichet Ruenchit, Siriraj Integrative Center for Neglected Parasitic Diseases, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand, E-mails: sirirat.wij@mahidol.ac.th, kedsara.pan@mahidol.ac.th, kosol.run@mahidol.ac.th, and pichet.rue@mahidol.edu. Sukhum Silpa-Archa, Department of Ophthalmology, Rajavithi Hospital, College of Medicine, Rangsit University, Bangkok, Thailand, E-mail: sukhumsilp@gmail.com. Matya Suwansirikul, Buddhasothorn Hospital, Ministry of Public Health, Na Mueang, Thailand, E-mail: s_matya@hotmail.com. Peerasak Chortrakarnkij, Division of Plastic and Reconstructive Surgery, Department of Surgery, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand, E-mail: peerasak.cho@mahidol.ac.th. Kanok Preativatanyou, Center of Excellence in Vector Biology and Vector-Borne Disease, Chulalongkorn University, Bangkok, Thailand, and Department of Parasitology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand, E-mail: kanok.pr@chula.ac.th. Sirichit Wongkamchai, Siriraj Integrative Center for Neglected Parasitic Diseases, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand, E-mail: sirichit.won@mahidol.ac.th.
- ABSTRACT.
- INTRODUCTION
- MATERIALS AND METHODS
- RESULTS
- Clinical and histopathological data of atypical filariasis cases.
- Molecular characterization and phylogenetic analysis of filarial mt 12 S rRNA gene.
- Molecular characterization and phylogenetic analysis of filarial COI.
- Molecular characterization and phylogenetic analysis of filarial ITS1.
- Molecular detection and phylogenetic analysis of Wolbachia endosymbionts.
- DISCUSSION
- Supplemental Materials
- ACKNOWLEDGMENT
- REFERENCES
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Whitworth JA , Hewitt K , 2005. Filariasis. Medicine (Baltimore) 33: 61–64.
-
2.
Centers for Disease Control and Prevention , 2019. Lymphatic Filariasis. Available at: https://www.cdc.gov/filarial-worms/about/lymphatic-filariasis.html. Accessed April 18, 2019.
-
3.
Simonsen PE , Fischer PU , Hoerauf A , Weil GJ , 2009. The filariases. Farrar J, Hotez PJ, Junghanss T, Kang G, Lalloo D, White NJ, eds. Manson’s Tropical Diseases, 23rd ed. Philadelphia, PA: Saunders Ltd., 737–765.
-
4.
Orihel TC , Eberhard ML , 1998. Zoonotic filariasis. Clin Microbiol Rev 11: 366.
-
5.
Dreyer G , Dreyer P , Piessens W , 1999. Extralymphatic disease due to bancroftian filariasis. Braz J Med Biol Res 32: 1467–1472.
-
6.
Gutierrez Y , 2000. Diagnostic Pathology of Parasitic Infections with Clinical Correlations, 2nd ed. New York, NY: Oxford University Press.
-
7.
Vaid SJ , Luthra A , Karnik S , Ahuja AT , 2011. Facial wrigglies: Live extralymphatic filarial infestation in subcutaneous tissues of the head and neck. Br J Radiol 84: e126–e129.
-
8.
Simón F , González-Miguel J , Diosdado A , Gómez PJ , Morchón R , Kartashev V , 2017. The complexity of zoonotic filariasis episystem and its consequences: A multidisciplinary view. BioMed Res Int 2017: 6436130.
-
9.
Somsap Y , Boonroumkaew P , Somsap A , Rodpai R , Sadaow L , Sanpool O , Maleewong W , Intapan P , 2021. Ocular dirofilariasis case in Thailand confirmed by molecular analysis to be caused by Dirofilaria immitis. Am J Trop Med Hyg 106: 204.
-
10.
Sukudom P , Phumee A , Siriyasatien P , 2018. First report on subconjunctival dirofilariasis in Thailand caused by a Dirofilaria sp. closely related to D. hongkongenesis. Acad J Sci Res 6: 114–116.
-
11.
Nunthanid P , Roongruanchai K , Wongkamchai S , Sarasombath PT , 2020. Case report: Periorbital filariasis caused by Brugia malayi. Am J Trop Med Hyg 103: 2336–2338.
-
12.
Thongpiya J , Sa-nguanraksa D , Samarnthai N , Sarasombath PT , 2020. Filariasis of the breast caused by Brugia pahangi: A concomitant finding with invasive ductal carcinoma. Parasitol Int 80: 102203.
-
13.
Thongpiya J , Kreetitamrong S , Thongsit T , Toothong T , Rojanapanus S , Sarasombath P , 2021. The first case report of subcutaneous dirofilariasis caused by Dirofilaria repens in Thailand. Trop Parasitol 11: 125–127.
-
14.
Suphap N , Somkijrungroj T , Kongwattananon W , Supawatjariyakul W , Pataradool T , Kraivichian K , Jantarabenjakul W , Tulvatana W , Preativatanyou K , 2024. Ocular Brugia pahangi filariasis complicated by severe macular damage in Thailand: Case report and literature review. Am J Trop Med Hyg 110: 1158–1164.
-
15.
Taylor MJ , 2000. Wolbachia bacteria of filarial nematodes in the pathogenesis of disease and as a target for control. Trans R Soc Trop Med Hyg 94: 596–598.
-
16.
Bouchery T , Karadjian G , Nieguitsila A , Martin C , 2013. The symbiotic role of Wolbachia in Onchocercidae and its impact on filariasis. Clin Microbiol Infect 19: 131–140.
-
17.
Wongkamchai S , Monkong N , Mahannol P , Taweethavonsawat P , Loymak S , Foongladda S , 2013. Rapid detection and identification of Brugia Malayi, B. Pahangi, and Dirofilaria immitis by high-resolution melting assay. Vector Borne Zoonotic Dis 13: 31–36.
-
18.
Casiraghi M , Favia G , Cancrini G , Bartoloni A , Bandi C , 2001. Molecular identification of Wolbachia from the filarial nematode Mansonella ozzardi. Parasitol Res 87: 417–420.
-
19.
Nuchprayoon S , Junpee A , Nithiuthai S , Chungpivat S , Suvannadabba S , Poovorawan Y , 2006. Detection of filarial parasites in domestic cats by PCR-RFLP of ITS1. Vet Parasitol 140: 366–372.
-
20.
Werren JH , Windsor DM , 2000. Wolbachia infection frequencies in insects: Evidence of a global equilibrium? Proc Biol Sci 267: 1277–1285.
-
21.
Zhou W , Rousset F , O’Neil S , 1998. Phylogeny and PCR-based classification of Wolbachia strains using wsp gene sequences. Proc Biol Sci 265: 509–515.
-
22.
Camacho C , Boratyn GM , Joukov V , Vera Alvarez R , Madden TL , 2023. ElasticBLAST: Accelerating sequence search via cloud computing. BMC Bioinformatics 24: 117.
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