Journal of Kerman University of Medical Sciences

Molecular Analysis of the Presence of Wolbachia in Indigenous Human Louse Populations in Iran

Document Type : Original Article

Authors

Samira Firooziyan 1, 2
Mohammad Ali Oshaghia 3
Saber Gholizadeh 1, 2

1 Pathogens and Vectors Research Center, Cellular and Molecular Medicine Research Institute, Urmia University of Medical Sciences, Urmia, Iran

2 Department of Medical Entomology and Vector Control, School of Public Health, Urmia University of Medical Sciences, Urmia, Iran

3 Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

https://doi.org/10.34172/jkmu.4093
Abstract
Introduction: Human lice infestation continues to be a public health challenge. Head lice not only cause physical irritation, but also lead to social and psychological problems. Body lice are vectors of several infectious diseases. While chemical control remains the mainstay of management, increasing insecticide resistance highlights the need for alternative strategies. Investigating endosymbiotic bacteria such as Wolbachia may provide new ways to control lice populations.
Methods: Samples were collected from volunteers in four provinces of Iran. DNA extraction was performed according to the manufacturer’s instructions with minor adjustments to improve results. The presence of Wolbachia was assessed using PCR and nested-PCR methods targeting the Wolbachia surface protein (wsp), glutamyl-tRNA amidotransferase B subunit (gatB), and cytochrome oxidase subunit 1 (coxA) genes. DNA isolated from Wolbachia-infected Culex mosquitoes was used as a positive control to confirm the effectiveness of the assay. Distilled deionized water was also used as a negative control to ensure no contamination or false positives.
Results: No Wolbachia DNA was found in any of the lice samples when the results were compared to the positive control. Amplification using the wsp, gatB, and coxA primer sets did not show any positive results in PCR or nested-PCR assays.
Conclusion: The absence of detectable Wolbachia in the louse samples of this study may be due to genetic variation in the primer binding regions or low bacterial density. Further molecular investigations using other markers and optimized primers are recommended to clarify the possible presence of Wolbachia in Iranian louse populations.

Keywords

Pediculus
Wolbachia
Iran
Subjects
  1. Badiaga S, Brouqui P. Human louse-transmitted infectious diseases. Clin Microbiol Infect 2012;18(4):332-7. doi:10.1111/ j.1469-0691.2012.03778.x
  2. Firooziyan S, Sadaghianifar A, Taghilou B, Galavani H, Ghaffari E, Gholizadeh S. Identification of novel voltage-gated sodium channel mutations in human head and body lice (Phthiraptera: Pediculidae). J Med Entomol 2017;54(5):1337- 43. doi:10.1093/jme/tjx107
  3. Aliverdi F, Mahmoodi Z, Mehdizadeh Tourzani Z, Amini L, Mohamadi F. The relationship between social network use and students’ quality of life. J Kerman Univ Med Sci 2024;31(6):299-304. doi:10.34172/jkmu.2024.46
  4. Mumcuoglu KY. Prevention and treatment of head lice in children. Paediatr Drugs 1999;1(3):211-8. doi:10.2165/00128072-199901030-00005
  5. Clark JM. Determination, mechanism and monitoring of knockdown resistance in permethrin-resistant human head lice, Pediculus humanus capitis. J Asia Pac Entomol 2009;12(1):1-7. doi:10.1016/j.aspen.2008.10.007
  6. Burkhart CG, Burkhart CN. Head lice therapies revisited. Dermatol Online J 2006;12(6):3. doi:10.5070/d35dz638vq
  7. Burgess IF, Burgess NA. Dimeticone 4% liquid gel found to kill all lice and eggs with a single 15 minute application. BMC Res Notes 2011;4:15. doi:10.1186/1756-0500-4-15
  8. Kurt O, Balcioğlu IC, Burgess IF, Limoncu ME, Girginkardeşler N, Tabak T, et al. Treatment of head lice with dimeticone 4% lotion: comparison of two formulations in a randomised controlled trial in rural Turkey. BMC Public Health 2009;9:441. doi:10.1186/1471-2458-9-441
  9. Kamalinejad M, Sharif A, Gachkar L, Ehsani A, Ahmadian-Atta M, Torabi MR, et al. Investigating the efficacy of sumac topical solution against permethrin-resistant human head lice. Int J Med Toxicol Forensic Med 2019;9(3):117-24. doi: 10.32598/ ijmtfm.v9i3.25296.
  10. Sazama EJ, Ouellette SP, Wesner JS. Bacterial endosymbionts are common among, but not necessarily within, insect species. Environ Entomol 2019;48(1):127-33. doi:10.1093/ee/nvy188
  11. O’Neill SL, Giordano R, Colbert AM, Karr TL, Robertson HM. 16S rRNA phylogenetic analysis of the bacterial endosymbionts associated with cytoplasmic incompatibility in insects. Proc Natl Acad Sci U S A 1992;89(7):2699-702. doi:10.1073/pnas.89.7.2699
  12. Rousset F, Bouchon D, Pintureau B, Juchault P, Solignac M. Wolbachia endosymbionts responsible for various alterations of sexuality in arthropods. Proc Biol Sci 1992;250(1328):91-8. doi:10.1098/rspb.1992.0135
  13. Stouthamer R, Breeuwert JA, Luck RF, Werren JH. Molecular identification of microorganisms associated with parthenogenesis. Nature 1993;361(6407):66-8. doi:10.1038/361066a0
  14. Simsek S, Ciftci AT. Serological and molecular detection of Dirofilaria species in stray dogs and investigation of Wolbachia DNA by PCR in Turkey. J Arthropod Borne Dis 2016;10(4):445-53.
  15. Sazama EJ, Bosch MJ, Shouldis CS, Ouellette SP, Wesner JS. Incidence of Wolbachia in aquatic insects. Ecol Evol 2017;7(4):1165-9. doi:10.1002/ece3.2742
  16. Werren JH, Windsor D, Guo LR. Distribution of Wolbachia among neotropical arthropods. Proc R Soc Lond B Biol Sci 1995;262(1364):197-204. doi:10.1098/rspb.1995.0196
  17. Kozek WJ, Rao RU. The discovery of Wolbachia in arthropods and nematodes–a historical perspective. In: Hoerauf A, Rao RU, eds. Wolbachia: A Bug’s Life in Another Bug. Karger Medical and Scientific Publishers; 2007. p. 1-14. doi:10.1159/000104228
  18. Werren JH, Windsor DM. Wolbachia infection frequencies in insects: evidence of a global equilibrium? Proc Biol Sci 2000;267(1450):1277-85. doi:10.1098/rspb.2000.1139
  19. Kittayapong P, Jamnongluk W, Thipaksorn A, Milne JR, Sindhusake C. Wolbachia infection complexity among insects in the tropical rice-field community. Mol Ecol 2003;12(4):1049-60. doi:10.1046/j.1365-294x.2003.01793.x
  20. Werren JH, Zhang W, Guo LR. Evolution and phylogeny of Wolbachia: reproductive parasites of arthropods. Proc Biol Sci 1995;261(1360):55-63. doi:10.1098/rspb.1995.0117
  21. Bensaadi-Merchermek N, Salvado JC, Cagnon C, Karama S, Mouchès C. Characterization of the unlinked 16S rDNA and 23S-5S rRNA operon of Wolbachia pipientis, a prokaryotic parasite of insect gonads. Gene 1995;165(1):81- 6. doi:10.1016/0378-1119(95)00539-i
  22. Kirkness EF, Haas BJ, Sun W, Braig HR, Perotti MA, Clark JM, et al. Genome sequences of the human body louse and its primary endosymbiont provide insights into the permanent parasitic lifestyle. Proc Natl Acad Sci U S A 2010;107(27):12168-73. doi:10.1073/pnas.1003379107
  23. Werren JH. Wolbachia and speciation. In: Endless Forms: Species and Speciation. Oxford University Press; 1998. p. 245-60. doi:10.1093/oso/9780195109009.003.0018
  24. Lo N, Casiraghi M, Salati E, Bazzocchi C, Bandi C. How many Wolbachia supergroups exist? Mol Biol Evol 2002;19(3):341- 6. doi:10.1093/oxfordjournals.molbev.a004087
  25. Bordenstein S, Rosengaus RB. Discovery of a novel Wolbachia super group in Isoptera. Curr Microbiol 2005;51(6):393-8. doi:10.1007/s00284-005-0084-0
  26. Lo N, Paraskevopoulos C, Bourtzis K, O’Neill SL, Werren JH, Bordenstein SR, et al. Taxonomic status of the intracellular bacterium Wolbachia pipientis. Int J Syst Evol Microbiol 2007;57(Pt 3):654-7. doi:10.1099/ijs.0.64515-0
  27. Ros VI, Fleming VM, Feil EJ, Breeuwer JA. How diverse is the genus Wolbachia? Multiple-gene sequencing reveals a putatively new Wolbachia supergroup recovered from spider mites (Acari: Tetranychidae). Appl Environ Microbiol 2009;75(4):1036-43. doi:10.1128/aem.01109-08
  28. Covacin C, Barker SC. Supergroup F Wolbachia bacteria parasitise lice (Insecta: Phthiraptera). Parasitol Res 2007;100(3):479-85. doi:10.1007/s00436-006-0309-6
  29. Zhou W, Rousset F, O’Neil S. Phylogeny and PCR-based classification of Wolbachia strains using wsp gene sequences. Proc Biol Sci 1998;265(1395):509-15. doi:10.1098/ rspb.1998.0324
  30. Baldo L, Desjardins CA, Russell JA, Stahlhut JK, Werren JH. Accelerated microevolution in an outer membrane protein (OMP) of the intracellular bacteria Wolbachia. BMC Evol Biol 2010;10:48. doi:10.1186/1471-2148-10-48
  31. Anderson M. Detection of Wolbachia in Human Lice. GS4 Georgia Southern Student Scholars Symposium. Georgia Southern University; 2016. doi:10.20429/gs4.2016.009
  32. Bleidorn C, Gerth M. A critical re-evaluation of multilocus sequence typing (MLST) efforts in Wolbachia. FEMS Microbiol. Ecol 2018;94(1):163. doi:10.1093/femsec/fix163
  33. Baldo L, Dunning Hotopp JC, Jolley KA, Bordenstein SR, Biber SA, Choudhury RR, et al. Multilocus sequence typing system for the endosymbiont Wolbachia pipientis. Appl Environ Microbiol 2006;72(11):7098-110. doi:10.1128/aem.00731- 06
  34. Werren JH. Biology of Wolbachia. Annu Rev Entomol 1997;42:587-609. doi:10.1146/annurev.ento.42.1.587
  35. Bozorg-Omid F, Oshaghi MA, Vahedi M, Karimian F, Seyyed- Zadeh SJ, Chavshin AR. Wolbachia infection in West Nile virus vectors of northwest Iran. Appl Entomol Zool 2020;55(1):105- 13. doi:10.1007/s13355-019-00658-6
  36. Maleki-Ravasan N, Akhavan N, Raz A, Jafari M, Zakeri S, Dinparast Djadid N. Co-occurrence of pederin-producing and Wolbachia endobacteria in Paederus fuscipes Curtis, 1840 (Coleoptera: Staphilinidae) and its evolutionary consequences. Microbiologyopen 2019;8(7):e00777. doi:10.1002/mbo3.777
  37. Karimi J, Darsouei R. Presence of the endosymbiont Wolbachia among some fruit flies (Diptera: Tephritidae) from Iran: a multilocus sequence typing approach. J Asia Pac Entomol 2014;17(1):105-12. doi:10.1016/j.aspen.2013.11.002
  38. Bordbar A, Soleimani S, Fardid F, Zolfaghari MR, Parvizi P. Three strains of Wolbachia pipientis and high rates of infection in Iranian sandfly species. Bull Entomol Res 2014;104(2):195- 202. doi:10.1017/s0007485313000631
  39. Allen JM, Reed DL, Perotti MA, Braig HR. Evolutionary relationships of “Candidatus Riesia spp.,” endosymbiotic enterobacteriaceae living within hematophagous primate lice. Appl Environ Microbiol 2007;73(5):1659-64. doi:10.1128/ aem.01877-06
  40. Sasaki-Fukatsu K, Koga R, Nikoh N, Yoshizawa K, Kasai S, Mihara M, et al. Symbiotic bacteria associated with stomach discs of human lice. Appl Environ Microbiol 2006;72(11):7349-52. doi:10.1128/aem.01429-06
  41. Perotti MA, Kirkness EF, Reed DL, Braig HR. Endosymbionts of lice. In: Insect Symbiosis. Vol 3. CRC Press; 2008. p. 205-19. doi:10.1201/9781420064117.ch9
  42. Kyei-Poku GK, Colwell DD, Coghlin P, Benkel B, Floate KD. On the ubiquity and phylogeny of Wolbachia in lice. Mol Ecol 2005;14(1):285-94. doi:10.1111/j.1365-294X.2004.02409.x
  43. Veracx A, Raoult D. Biology and genetics of human head and body lice. Trends Parasitol 2012;28(12):563-71. doi:10.1016/j.pt.2012.09.003
  44. Kittayapong P, Baisley KJ, Baimai V, O’Neill SL. Distribution and diversity of Wolbachia infections in Southeast Asian mosquitoes (Diptera: Culicidae). J Med Entomol 2000;37(3):340-5. doi:10.1093/jmedent/37.3.340
  45. Osei-Poku J, Han C, Mbogo CM, Jiggins FM. Identification of Wolbachia strains in mosquito disease vectors. PLoS One 2012;7(11):e49922. doi:10.1371/journal.pone.0049922
  46. Johnson KN. The impact of Wolbachia on virus infection in mosquitoes. Viruses 2015;7(11):5705-17. doi:10.3390/ v7112903
  47. Schneider DI, Klasson L, Lind AE, Miller WJ. More than fishing in the dark: PCR of a dispersed sequence produces simple but ultrasensitive Wolbachia detection. BMC Microbiol 2014;14:121. doi:10.1186/1471-2180-14-121

 

Journal of Kerman University of Medical Sciences
Volume 33
Pages 1-5

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