Morphological and Molecular Analysis of Enterobius vermicularis with Syphacia spp.
-
Maliheh Mostafazadeh
,
Fattaneh Mikaeili
,
Aref Teimouri
,
Seyed Mahmoud Sadjjadi
,
Yosef Sharifi
,
Manouchehr Shirani
,
Amin Shamsaei Sarvestani
,
Meysam Sharifdini
Abstract
Background: Syphacia (rodent pinworm), a common nematode in the colon of rodents, has rarely been reported in humans. The morphological identification of some pinworm species is difficult, especially in those cases where only fragments of worms are recovered. We aimed to identify isolates of Syphacia spp. in rodents using morphological and molecular approaches, and to compare them with human Enterobius vermicularis.
Methods: This study was carried out on 10 adult Syphacia worms collected from BALB/c mice and five adult Enterobius vermicularis from humans. Morphological features using existing keys and PCR-sequencing of ITS1 and 5.8s regions were applied to identify Syphacia spp., and its comparison with E. vermicularis.
Results: The total length of esophagus, the length and width of esophageal bulb, and the length and width of the right and left cephalic alae of E. vermicularis were larger than Syphacia. These parameters demonstrated significant morphological differences between E. vermicularis and Syphacia. PCR successfully produced amplicons of approximately 414 bp for Syphacia and 473 bp for E. vermicularis. The molecular method identified the Oxyurid nematodes isolated from BALB/c mice as Syphacia obvelata. The pairwise comparison revealed no differences in nucleotide sequences among S. obvelata isolates, and the sequences were identical and exhibited 100% homology.
Conclusions: This study demonstrated genetic and morphological differences between E. vermicularis and S. obvelata. Since laboratory mice (Mus musculus) are the specific hosts for S. obvelata, controlling these animals is critical to maintaining public health.
1. Baker D. Natural pathogens of laboratory mice, rats, and rabbits and their effects on research. Clin Microbiol Rev. 1998;11:231-66.
2. Kucik CJ, Martin GL, Sortor BV. Common intestinal parasites. Am Fam Physician. 2004;69(5):1161-8.
3. Sundar SB, Harikrishnan TJ, Ravi LB, et al. Prevalence of syphaciosis in laboratory rodents with emphasis on species identification, treatment and Th2 cytokine response. Indian J Anim Sci. 2018; 88:765-70.
4. Chan KF. Life-cycle studies on the nematode Syphacia obvelata. Am J Hyg. 1952; 56:14-21.
5. Muller R. Worms and Human Disease. 2nd ed. Cambridge (UK): CABI Publishing; 2002.
6. Meade TM, Watson J. Characterization of rat pinworm (Syphacia muris) epidemiology as a means to increase detection and elimination. J Am Assoc Lab Anim Sci. 2014;53:661-7.
7. Hamzavi Y, Khodayari MT, Davari A, et al. A systematic review and meta-analysis on prevalence of gastrointestinal helminthic infections in rodents of Iran: An emphasis on zoonotic aspects. Heliyon. 2024;10(11):e31955.
8. Lima VFS, Ramos RAN, Giannelli A, et al. Occurrence of zoonotic gastrointestinal parasites of rodents and the risk of human infection in different biomes of Brazil. Braz J Vet Med. 2021;43:e113820.
9. Riley WA. A mouse oxyurid, Syphacia obvelata, as a parasite of man. J Parasitol. 1919;6:89-93.
10. Dahmana H, Granjon L, Diagne C, et al. Rodents as Hosts of Pathogens and Related Zoonotic Disease Risk. Pathogens. 2020;9(3):202.
11. Jacoby RO, Fox JG, Davisson M. Biology and Diseases of Mice. In: Fox JG, Anderson LC, Loew FM, Quimby FW, editors. In American College of Laboratory Animal Medicine, Laboratory Animal Medicine (Second Edition). Academic Press; 2002, P. 35-120.
12. Bazzano T, Restel TI, Pinto RM, et al. Patterns of infection with nematodes Syphacia obvelata and Aspicularis tetraptera in conventionally maintained laboratory mice. Mem Inst Oswaldo Cruz. 2002;97:847-53.
13. Semenova SK, Romanova EA, Pyskov AP. Genetic differentiation of helminthes on the basis of data of polymerase chain reaction using random primers. Genetika. 1996;32(2):304-9.
14. Robles MR, Navone GT. A new species of Syphacia (Nematoda: Oxyuridae) from Oligoryzomys nigripes (Rodentia: Cricetidae) in Argentina. Parasitol Res. 2007;101(4):1069-75.
15. Parel JD, Galula JU, Ooi HK. Characterization of rDNA sequences from Syphacia obvelata, Syphacia muris, and Aspiculuris tetraptera and development of a PCR-based method for identification. Vet Parasitol. 2008;153(3-4):379-83.
16. Mc-Manus DP, Bowles J. Molecular genetic approaches to parasite identification: their value in diagnostic parasitology and systematics. Int J Parasitol. 1996;26:687-704.
17. Nakano T, Okamoto M, Ikeda Y, et al. Mitochondrial cytochrome c oxidase subunit 1 gene and nuclear rDNA regions of Enterobius vermicularis parasitic in captive chimpanzees with special reference to its relationship with pinworms in humans. Parasitol Res. 2006;100:51-7.
18. Okamoto M, Urushima H, Iwasa M, et al. Phylogenetic relationships of rodent pinworms (genus Syphacia) in Japan inferred from mitochondrial CO1 gene sequences. J Vet Med Sci. 2007;69(5):545-7.
19. Okamoto M, Urushima H, Hasegawa H. Phylogenetic relationships of rodent pinworms (genus Syphacia) in Japan inferred from 28S rDNA sequences. Parasitol Int. 2009;58:330-3.
20. Abdel-Gaber R. Syphacia obvelata (Nematode, Oxyuridae) infecting laboratory mice Mus musculus (Rodentia, Muridae): phylogeny and host-parasite relationship. Parasitol Res. 2016;115:975-85.
21. Kubiak K, Dzika E, Lukasz P. Enterobiasis epidemiology and molecular characterization of Enterobius vermicularis in healthy children in north-eastern Poland. Helminthologia. 2017;54(4):10.1515/helm-2017-0042.
22. Julius RS, Schwan EV, Chimimba CT. Molecular characterization of cosmopolitan and potentially co-invasive helminths of commensal, murid rodents in Gauteng Province, South Africa. Parasitol Res. 2018;117(6):1729-36.
23. Stewart A, Lowe A, Smales L, et al. Parasitic nematodes of the genus Syphacia Seurat, 1916 infecting Muridae in the British Isles, and the peculiar case of Syphacia frederici. Parasitology. 2018;145(3):269-80.
24. Tavan A, Mikaeili F, Sadjjadi SM, et al. Prevalence and genotype distribution of Enterobius vermicularis among kindergarteners in Shiraz and Khorramabad cities, Iran. Asian Pac J Trop Med. 2020;13(7):308-13.
25. Tomanakan K, Sanpool O, Chamavit P, et al. Genetic variation of Enterobius vermicularis among schoolchildren in Thailand. J Helminthol. 2020;94:e7.
26. Gorelysheva DI, Gromov AR, Lavrenchenko LA, et al. Genetic diversity of Syphacia Seurat, 1916 (Nematoda: Oxyuridae) across the hybrid zone of their rodent hosts in Russia. Parasitol Res. 2021;120(6):2017-30.
27. Haghshenas M, Koosha M, Latifi A, et al. Detection of Enterobius vermicularis in archived formalin-fixed paraffin-embedded (FFPE) appendectomy blocks: It's potential to compare genetic variations based on mitochondrial DNA (cox1) gene. PLoS One. 2023;18(2):e0281622.
28. Tanideh N, Sadjjadi S, Mohammadzadeh T, et al. Helminthic infections of laboratory animals in animal house of Shiraz University of Medical Sciences and the potential risks of zoonotic infections for researchers. Iran Red Crescent Med J. 2010;12(2):151-7.
29. Zahabiun F, Sadjjadi SM, Esfandiari F. Development of a Double Glass Mounting Method Using Formaldehyde Alcohol Azocarmine Lactophenol (FAAL) and its Evaluation for Permanent Mounting of Small Nematodes. Iran J Parasitol. 2015;10(4):617-24.
30. Feldman SH, Bowman SG. Molecular phylogeny of the pinworms of mice, rats and rabbits, and its use to develop molecular beacon assays for the detection of pinworms in mice. Lab Anim (NY). 2007;36:43-50.
31. Kiene F, Andriatsitohaina B, Ramsay MS, et al. Habitat fragmentation and vegetation structure impact gastrointestinal parasites of small mammalian hosts in Madagascar. Ecol Evol. 2021;11(11):6766-88.
32. Zelck UE, Bialek R, Weiss M. Molecular phylogenetic analysis of Enterobius vermicularis and development of an 18S ribosomal DNA-targeted diagnostic PCR. J Clin Microbiol. 2011;49(4):1602-4.
33. Whary MT, Baumgarth N, Fox JG, Barthold SW. Biology and Diseases of Mice. Fox JG, Anderson LC, Otto GM, Pritchett-Corning KR, Whary MT, editors. In American College of Laboratory Animal Medicine, Laboratory Animal Medicine (Third Edition). Academic Press; 2015. P. 43-149.
34. Patel B, Sharma T, Bhatt GC, et al. Enterobius vermicularis: An unusual cause of recurrent urinary tract infestation in a 7-year-old girl: Case report and review of the literature. Trop Doct. 2015;45(2):132-4.
35. Chhetri S, Al Mamari AH, Al Awfi MM, et al. Enterobius vermicularis Related Acute Appendicitis: A Case Report and Review of the Literature. Infect Dis Rep. 2023;15(4):417-24.
36. Pinto RM, Vicente JJ, Noronha D, et al. Helminth parasites of conventionally maintained laboratory mice. Mem Inst Oswaldo Cruz. 1994;89(1):33-40.
37. Landaeta-Aqueveque CA, Robles MDR, Cattan PE. The community of gastrointestinal helminths in the house mouse, Mus musculus, in Santiago, Chile. Parasitol Latinoam. 2007;62:165-9.
38. Hussey K. Syphacia muris vs. Syphacia obvelata in laboratory rats and mice. J Parasitol. 1957;43:555-9.
39. Khalil AI, Lashein GH, MorsyGH, et al. Oxyurids of wild and laboratory rodents from Egypt. Life Sci J. 2014;11(3):94-107.
40. Wu ML, Kuksuk LK, Olinger EJ. Enterobius vermicularis. Arch Pathol Lab Med. 2000;124(4):647-8.
41. Sharifdini M, Heckmann RA, Mikaeili F. The morphological and molecular characterization of Baylisascaris devosi Sprent, 1952 (Ascaridoidea, Nematoda), collected from Pine marten (Martes martes) in Iran. Parasit Vectors. 2021;14(1):33.
42. Shafiei R, Sarkari B, Moshfe A. A Consistent PCR-RFLP Assay Based on ITS-2 Ribosomal DNA for Differentiation of Fasciola Species. Iran J Basic Med Sci. 2013;16:1266-69.
43. Mikaeili F, Mathis A, Deplazes P, et al. Differentiation of Toxocara canis and Toxocara cati based on PCR-RFLP analyses of rDNA-ITS and mitochondrial cox1 and nad1 regions. Acta Parasitol. 2017;62(3):549-56.
44. Mohammed OB, Al-Shaebi EM, Omer SA, et al. Morphological and molecular studies on a nematode parasite, Syphacia obvelata, infecting laboratory mice (Mus musculus) in Saudi Arabia. Arq Bras Med Vet Zootec. 2024;76:84-92.
45. Baird SJE, Ribas A, Macholan M, et al. Where are the wormy mice? A re-examination of hybrid parasitism in the European house mouse hybrid zone. Evolution. 2012;66(9):2757-72.
2. Kucik CJ, Martin GL, Sortor BV. Common intestinal parasites. Am Fam Physician. 2004;69(5):1161-8.
3. Sundar SB, Harikrishnan TJ, Ravi LB, et al. Prevalence of syphaciosis in laboratory rodents with emphasis on species identification, treatment and Th2 cytokine response. Indian J Anim Sci. 2018; 88:765-70.
4. Chan KF. Life-cycle studies on the nematode Syphacia obvelata. Am J Hyg. 1952; 56:14-21.
5. Muller R. Worms and Human Disease. 2nd ed. Cambridge (UK): CABI Publishing; 2002.
6. Meade TM, Watson J. Characterization of rat pinworm (Syphacia muris) epidemiology as a means to increase detection and elimination. J Am Assoc Lab Anim Sci. 2014;53:661-7.
7. Hamzavi Y, Khodayari MT, Davari A, et al. A systematic review and meta-analysis on prevalence of gastrointestinal helminthic infections in rodents of Iran: An emphasis on zoonotic aspects. Heliyon. 2024;10(11):e31955.
8. Lima VFS, Ramos RAN, Giannelli A, et al. Occurrence of zoonotic gastrointestinal parasites of rodents and the risk of human infection in different biomes of Brazil. Braz J Vet Med. 2021;43:e113820.
9. Riley WA. A mouse oxyurid, Syphacia obvelata, as a parasite of man. J Parasitol. 1919;6:89-93.
10. Dahmana H, Granjon L, Diagne C, et al. Rodents as Hosts of Pathogens and Related Zoonotic Disease Risk. Pathogens. 2020;9(3):202.
11. Jacoby RO, Fox JG, Davisson M. Biology and Diseases of Mice. In: Fox JG, Anderson LC, Loew FM, Quimby FW, editors. In American College of Laboratory Animal Medicine, Laboratory Animal Medicine (Second Edition). Academic Press; 2002, P. 35-120.
12. Bazzano T, Restel TI, Pinto RM, et al. Patterns of infection with nematodes Syphacia obvelata and Aspicularis tetraptera in conventionally maintained laboratory mice. Mem Inst Oswaldo Cruz. 2002;97:847-53.
13. Semenova SK, Romanova EA, Pyskov AP. Genetic differentiation of helminthes on the basis of data of polymerase chain reaction using random primers. Genetika. 1996;32(2):304-9.
14. Robles MR, Navone GT. A new species of Syphacia (Nematoda: Oxyuridae) from Oligoryzomys nigripes (Rodentia: Cricetidae) in Argentina. Parasitol Res. 2007;101(4):1069-75.
15. Parel JD, Galula JU, Ooi HK. Characterization of rDNA sequences from Syphacia obvelata, Syphacia muris, and Aspiculuris tetraptera and development of a PCR-based method for identification. Vet Parasitol. 2008;153(3-4):379-83.
16. Mc-Manus DP, Bowles J. Molecular genetic approaches to parasite identification: their value in diagnostic parasitology and systematics. Int J Parasitol. 1996;26:687-704.
17. Nakano T, Okamoto M, Ikeda Y, et al. Mitochondrial cytochrome c oxidase subunit 1 gene and nuclear rDNA regions of Enterobius vermicularis parasitic in captive chimpanzees with special reference to its relationship with pinworms in humans. Parasitol Res. 2006;100:51-7.
18. Okamoto M, Urushima H, Iwasa M, et al. Phylogenetic relationships of rodent pinworms (genus Syphacia) in Japan inferred from mitochondrial CO1 gene sequences. J Vet Med Sci. 2007;69(5):545-7.
19. Okamoto M, Urushima H, Hasegawa H. Phylogenetic relationships of rodent pinworms (genus Syphacia) in Japan inferred from 28S rDNA sequences. Parasitol Int. 2009;58:330-3.
20. Abdel-Gaber R. Syphacia obvelata (Nematode, Oxyuridae) infecting laboratory mice Mus musculus (Rodentia, Muridae): phylogeny and host-parasite relationship. Parasitol Res. 2016;115:975-85.
21. Kubiak K, Dzika E, Lukasz P. Enterobiasis epidemiology and molecular characterization of Enterobius vermicularis in healthy children in north-eastern Poland. Helminthologia. 2017;54(4):10.1515/helm-2017-0042.
22. Julius RS, Schwan EV, Chimimba CT. Molecular characterization of cosmopolitan and potentially co-invasive helminths of commensal, murid rodents in Gauteng Province, South Africa. Parasitol Res. 2018;117(6):1729-36.
23. Stewart A, Lowe A, Smales L, et al. Parasitic nematodes of the genus Syphacia Seurat, 1916 infecting Muridae in the British Isles, and the peculiar case of Syphacia frederici. Parasitology. 2018;145(3):269-80.
24. Tavan A, Mikaeili F, Sadjjadi SM, et al. Prevalence and genotype distribution of Enterobius vermicularis among kindergarteners in Shiraz and Khorramabad cities, Iran. Asian Pac J Trop Med. 2020;13(7):308-13.
25. Tomanakan K, Sanpool O, Chamavit P, et al. Genetic variation of Enterobius vermicularis among schoolchildren in Thailand. J Helminthol. 2020;94:e7.
26. Gorelysheva DI, Gromov AR, Lavrenchenko LA, et al. Genetic diversity of Syphacia Seurat, 1916 (Nematoda: Oxyuridae) across the hybrid zone of their rodent hosts in Russia. Parasitol Res. 2021;120(6):2017-30.
27. Haghshenas M, Koosha M, Latifi A, et al. Detection of Enterobius vermicularis in archived formalin-fixed paraffin-embedded (FFPE) appendectomy blocks: It's potential to compare genetic variations based on mitochondrial DNA (cox1) gene. PLoS One. 2023;18(2):e0281622.
28. Tanideh N, Sadjjadi S, Mohammadzadeh T, et al. Helminthic infections of laboratory animals in animal house of Shiraz University of Medical Sciences and the potential risks of zoonotic infections for researchers. Iran Red Crescent Med J. 2010;12(2):151-7.
29. Zahabiun F, Sadjjadi SM, Esfandiari F. Development of a Double Glass Mounting Method Using Formaldehyde Alcohol Azocarmine Lactophenol (FAAL) and its Evaluation for Permanent Mounting of Small Nematodes. Iran J Parasitol. 2015;10(4):617-24.
30. Feldman SH, Bowman SG. Molecular phylogeny of the pinworms of mice, rats and rabbits, and its use to develop molecular beacon assays for the detection of pinworms in mice. Lab Anim (NY). 2007;36:43-50.
31. Kiene F, Andriatsitohaina B, Ramsay MS, et al. Habitat fragmentation and vegetation structure impact gastrointestinal parasites of small mammalian hosts in Madagascar. Ecol Evol. 2021;11(11):6766-88.
32. Zelck UE, Bialek R, Weiss M. Molecular phylogenetic analysis of Enterobius vermicularis and development of an 18S ribosomal DNA-targeted diagnostic PCR. J Clin Microbiol. 2011;49(4):1602-4.
33. Whary MT, Baumgarth N, Fox JG, Barthold SW. Biology and Diseases of Mice. Fox JG, Anderson LC, Otto GM, Pritchett-Corning KR, Whary MT, editors. In American College of Laboratory Animal Medicine, Laboratory Animal Medicine (Third Edition). Academic Press; 2015. P. 43-149.
34. Patel B, Sharma T, Bhatt GC, et al. Enterobius vermicularis: An unusual cause of recurrent urinary tract infestation in a 7-year-old girl: Case report and review of the literature. Trop Doct. 2015;45(2):132-4.
35. Chhetri S, Al Mamari AH, Al Awfi MM, et al. Enterobius vermicularis Related Acute Appendicitis: A Case Report and Review of the Literature. Infect Dis Rep. 2023;15(4):417-24.
36. Pinto RM, Vicente JJ, Noronha D, et al. Helminth parasites of conventionally maintained laboratory mice. Mem Inst Oswaldo Cruz. 1994;89(1):33-40.
37. Landaeta-Aqueveque CA, Robles MDR, Cattan PE. The community of gastrointestinal helminths in the house mouse, Mus musculus, in Santiago, Chile. Parasitol Latinoam. 2007;62:165-9.
38. Hussey K. Syphacia muris vs. Syphacia obvelata in laboratory rats and mice. J Parasitol. 1957;43:555-9.
39. Khalil AI, Lashein GH, MorsyGH, et al. Oxyurids of wild and laboratory rodents from Egypt. Life Sci J. 2014;11(3):94-107.
40. Wu ML, Kuksuk LK, Olinger EJ. Enterobius vermicularis. Arch Pathol Lab Med. 2000;124(4):647-8.
41. Sharifdini M, Heckmann RA, Mikaeili F. The morphological and molecular characterization of Baylisascaris devosi Sprent, 1952 (Ascaridoidea, Nematoda), collected from Pine marten (Martes martes) in Iran. Parasit Vectors. 2021;14(1):33.
42. Shafiei R, Sarkari B, Moshfe A. A Consistent PCR-RFLP Assay Based on ITS-2 Ribosomal DNA for Differentiation of Fasciola Species. Iran J Basic Med Sci. 2013;16:1266-69.
43. Mikaeili F, Mathis A, Deplazes P, et al. Differentiation of Toxocara canis and Toxocara cati based on PCR-RFLP analyses of rDNA-ITS and mitochondrial cox1 and nad1 regions. Acta Parasitol. 2017;62(3):549-56.
44. Mohammed OB, Al-Shaebi EM, Omer SA, et al. Morphological and molecular studies on a nematode parasite, Syphacia obvelata, infecting laboratory mice (Mus musculus) in Saudi Arabia. Arq Bras Med Vet Zootec. 2024;76:84-92.
45. Baird SJE, Ribas A, Macholan M, et al. Where are the wormy mice? A re-examination of hybrid parasitism in the European house mouse hybrid zone. Evolution. 2012;66(9):2757-72.
| Files | ||
| Issue | Vol 20 No 3 (2025) | |
| Section | Original Article(s) | |
| Keywords | ||
| Morphology Molecular analysis Syphacia spp. Enterobius vermicularis | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Mostafazadeh M, Mikaeili F, Teimouri A, Sadjjadi SM, Sharifi Y, Shirani M, Shamsaei Sarvestani A, Sharifdini M. Morphological and Molecular Analysis of Enterobius vermicularis with Syphacia spp. Iran J Parasitol. 2025;20(3):454-464.
