Experimental and Applied Acarology

, Volume 68, Issue 4, pp 519–538 | Cite as

Evidence of female sex pheromones and characterization of the cuticular lipids of unfed, adult male versus female blacklegged ticks, Ixodes scapularis

  • Ann L. Carr
  • Daniel E. Sonenshine
  • John B. StriderJr.
  • R. Michael Roe


Copulation in Ixodes scapularis involves physical contact between the male and female (on or off the host), male mounting of the female, insertion/maintenance of the male chelicerae in the female genital pore (initiates spermatophore production), and the transfer of the spermatophore by the male into the female genital pore. Bioassays determined that male mounting behavior/chelicerae insertion required direct contact with the female likely requiring non-volatile chemical cues with no evidence of a female volatile sex pheromone to attract males. Unfed virgin adult females and replete mated adult females elicited the highest rates of male chelicerae insertion with part fed virgin adult females exhibiting a much lower response. Whole body surface hexane extracts of unfed virgin adult females and males, separately analyzed by GC–MS, identified a number of novel tick surface associated compounds: fatty alcohols (1-hexadecanol and 1-heptanol), a fatty amide (erucylamid), aromatic hydrocarbons, a short chain alkene (1-heptene), and a carboxylic acid ester (5β-androstane). These compounds are discussed in terms of their potential role in female–male communication. The two most abundant fatty acid esters found were butyl palmitate and butyl stearate present in ratios that were sex specific. Only 6 n-saturated hydrocarbons were identified in I. scapularis ranging from 10 to 18 carbons.


Ticks Pheromones Chemical communication Reproduction Cuticle Cuticular lipids Blacklegged tick Ixodes scapularis Lyme disease 



This work was funded by grants to RMR and DES from NIH (1R21AI096268) and NSF (IOS-0949194) and from support to RMR from the NCSU Ag. Research Station. ALC was also supported in part by a Graduate Student Teaching Assistantship from the Department of Entomology at North Carolina State University.

Compliance with ethical standards

Conflicts of interest

The authors have no conflicts of interest in the publication of the results reported in this paper.

Ethical standard

Students conducting research on this project have received training in a Graduate School approved ethics course at NC State University, which complies to the NSF standards.

Human and animal rights

No human subjects were used in the research reported in this paper. All use of animals in this study was carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. The protocols were approved by the Old Dominion University Institutional Animal Care and Use Committee (#10-018 and #10-032) and are on file at the Office of Research, Old Dominion University, Norfolk, Virginia. Tranquilizers (Acepromazine) were administered to the animals prior to handling to minimize anxiety and/or discomfort. No animal work was conducted at NC State University.


  1. Allen SA, Phillips JD, Taylor D, Sonenshine DE (1988) Genital sex pheromones of ixodid ticks: evidence for the role of fatty acids from the anterior reproductive tract in mating of Dermacentor variabilis and Dermacentor andersoni. J Insect Physiol 34:315–323CrossRefGoogle Scholar
  2. Allen SA, Sonenshine DE, Burridge MJ (2002) Tick pheromones and uses thereof. United States Patent Office, patent no. 6331297Google Scholar
  3. Benoit JB, Lopez-Martinez G, Philips SA, Elnitsky MA, Yoder JA, Lee RE Jr, Denlinger DL (2008) The seabird tick, Ixodes uriae, uses uric acid in penguin guano as a kairomone and guanine in tick feces as an assembly pheromone on the Antarctic Peninsula. Polar Biol 31:1445–1451CrossRefGoogle Scholar
  4. Bjostad LB, Wold WA, Roelofs WL (1987) Pheromone biosynthesis in lepidotpterans: desaturation and chain shortening. In: Prestwich GD, Blomquist GJ (eds) Pheromone biochemistry. Academic Press, Orlando, pp 77–120Google Scholar
  5. Caputo B, Dani FR, Horne GL, Petrarca V, Turillazzi S, Coluzzi M, Priestmand A, della Torre A (2005) Identification and composition of cuticular hydrocarbons of the major Afrotropical malaria vector Anopheles gambiae s.s. (Diptera: Culicidae): analysis of sexual dimorphism and age related changes. J Mass Spectrom 40:1595–1604CrossRefPubMedGoogle Scholar
  6. Dani FR (2006) Cuticular lipids as semiochemicals in paper wasps and other social insects. Ann Zool Fennici 43:500–514Google Scholar
  7. De Loof A (2006) Ecdysteroids: the overlooked sex steroid of insects? Males: the black box. Insect Sci 13:325–338CrossRefGoogle Scholar
  8. Dobrotvorsky AK, Tkachev AV (1995) Evidence of a volatile sex pheromone in the unfed adult taiga tick Ixodes persulcatus Schulze. In: 2nd international conference on tick-borne pathogens at the host-vector interface: an agenda for research, proceedings and abstract, vol 1, pp 372–375Google Scholar
  9. Dusbábek FP, Simek EAP, Bouman R, Zahradnikova H, Zemek R (2001) Composition and effect of several semiochemicals in Ixodes ricinus L. (Acari: Ixodidae). In: Blaszak C, Bucsek A (eds) Stawonogi Pasozyty i Nosiciele. Lublin, Wydawnictwo KGM, pp 33–43Google Scholar
  10. Estrada-Peña A (1993) Climate and cuticular hydrocarbon variations in Rhipicephalus sanguineus ticks (Acari: Ixodidae). Parasitol Res 79:512–516CrossRefPubMedGoogle Scholar
  11. Estrada-Peña A, Estrada-Peña R, Peiro JM (1992) Differentiation of Rhipicephalus ticks (Acari, Ixodidae) by gas-chromatography of cuticular hydrocarbons. J Parasitol 78:982–993CrossRefPubMedGoogle Scholar
  12. Estrada-Peña A, Castellá J, Siuda K (1994) Cuticular hydrocarbon composition and pheromone variability in sympatric populations of Ixodes ricinus ticks from Poland. Exp Appl Acarol 18:247–263CrossRefPubMedGoogle Scholar
  13. Fourie LJ, de Jager T, Petney TN (1988) Prolonger or repeated copulation and male longevity in the tick Ixodes rubicundus. J Parasitol 74:609–612CrossRefPubMedGoogle Scholar
  14. Graf JF (1978) Ecology and ethology of Ixodes ricinus (Ixodidae). B Swiss Entomol Soc 50:241–253Google Scholar
  15. Haggart DA, Davis EE (1981) Neurons sensitive to 2,6-dichlorophenol on the tarsi of the tick Amblyomma americanum (Acari: Ixodidae). J Med Entomol 3:187–193CrossRefGoogle Scholar
  16. Hamilton JGC, Sonenshine DE, Lushby WR (1989) Cholesteryl oleate; mounting sex pheromone of the hard tick, Dermacentor variabilis. (Say) (Acari: Ixodidae). J Insect Physiol 35:873–879CrossRefGoogle Scholar
  17. Heethoff M (2012) Regeneration of complex oil-gland secretions and its importance for chemical defense in an oribatid mite. J Chem Ecol 38:1116–1123CrossRefPubMedGoogle Scholar
  18. Howard RW, Lord JC (2003) Cuticular lipids of the booklouse, Liposcelis bostrychophila: hydrocarbons, aldehydes, fatty acids, and fatty acid amides. J Chem Ecol 29:615–627CrossRefPubMedGoogle Scholar
  19. Kiszewski AE, Spielman A (2002) Preprandial inhibition of re-mating in Ixodes ticks (Acari: Ixodidae). J Med Entomol 39:847–853CrossRefPubMedGoogle Scholar
  20. Kiszewski AE, Matuschka F, Spielman A (2001) Mating strategies and spermiogenesis in ixodid ticks. Annu Rev Entomol 46:167–182CrossRefPubMedGoogle Scholar
  21. Lenoir A, Cuvillier-Hot V, Devers A, Christidès J, Montigny F (2012) Ant cuticles: a trap for atmospheric phthalate contaminants. Sci Total Environ 441:209–212CrossRefPubMedGoogle Scholar
  22. Moorhouse DE, Heath CG (1975) Parasitism of female ticks by males of the genus Ixodes. J Med Entomol 12:571–572CrossRefPubMedGoogle Scholar
  23. Mulenga A (2014) Molecular biology and physiology of chemical communication. In: Sonenshine DE, Roe RM (eds) Biology of ticks, vol I. Oxford University Press, New York, pp 368–397Google Scholar
  24. Osterkamp J, Wahl U, Schmalfuss G, Haas W (1999) Host odor recognition in two tick species coded in a blend of vertebrate volatiles. J Comp Physiol 185:59–67CrossRefGoogle Scholar
  25. Paine TD, Millar JG, Hanlon CC, Hwang JS (1999) Identification of semiochemicals associated with Jeffrey pine beetle, Dendroctonus jeffreyi. J Chem Ecol 25:433–453CrossRefGoogle Scholar
  26. Shimshoni JA, Erster O, Cuneah O, Soback S, Shkap V (2013) Cuticular fatty acid profile analysis of three Rhipicephalus tick species (Acari: Ixodidae). Exp Appl Acarol 61:481–489CrossRefPubMedGoogle Scholar
  27. Soares SF, Borges MLF (2012) Electrophysiological responses of the olfactory receptors of the tick Amblyomma cajennense (Acari: Ixodidae) to host-related and tick pheromone-related synthetic compounds. Acta Trop 124:192–198CrossRefPubMedGoogle Scholar
  28. Sonenshine DE (2006) Tick pheromones and their use in tick control. Annu Rev Entomol 51:557–580CrossRefPubMedGoogle Scholar
  29. Sonenshine DE, Adams T, Allan SA, McLaughlin J, Webster FX (2003) Chemical composition of some components of the arrestment pheromones of the black-legged tick, Ixodes scapularis (Acari: Ixodidae) and their use in tick control. Med Entomol 40:849–859CrossRefGoogle Scholar
  30. Tkachev AV, Dobrotvorsky AK, Vjalkov AI, Morozov SV (2000) Chemical composition of lipophylic compounds from the body surface of unfed adult Ixodes persulcatus ticks (Acari: Ixodidae). Exp Appl Acarol 24:145–158CrossRefPubMedGoogle Scholar
  31. Torto B, Obeng Ofori D, Njagi PGN, Hassanali A, Amiani A (1994) Aggregation pheromones systems of adult gregarious desert locust Schistocerca gregaria. J Chem Ecol 20:1749–1762CrossRefPubMedGoogle Scholar
  32. Treverrow NL, Stone BF, Cowie M (1977) Aggregation pheromone in two Australian hard ticks, Ixodes holocylus and Aponomma concolor. Experientia 33:680–682CrossRefPubMedGoogle Scholar
  33. Waladde SM, Rice MJ (1982) The sensory basis of tick feeding behavior. In: Obenchain FD, Galun R (eds) Physiology of ticks. Pergamon Press, Oxford, pp 71–118CrossRefGoogle Scholar
  34. Xiao YH, Zhang JX, Li SQ (2009) A two-component female produced pheromone of the spider Pholcus beijingensis. J Chem Ecol 35:769–778CrossRefPubMedGoogle Scholar
  35. Zemek R, Bouman EAP, Socha R, Dusbábek F (2002) The effect of feeding status on sexual attractiveness of Ixodes ricinus (Acari: Ixodidae) females. Exp Appl Acarol 27:137–149CrossRefGoogle Scholar
  36. Zhang J, Salcedo C, Fang Y, Zhang R, Zhang Z (2012) An overlooked component: (z)-9-tetradecenal as a sex pheromone in Helicoverpa armigera. J Insect Physiol 58:1209–1216CrossRefPubMedGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2016

Authors and Affiliations

  • Ann L. Carr
    • 1
  • Daniel E. Sonenshine
    • 2
  • John B. StriderJr.
    • 1
  • R. Michael Roe
    • 1
  1. 1.Department of EntomologyNorth Carolina State UniversityRaleighUSA
  2. 2.Department of Biological SciencesOld Dominion UniversityNorfolkUSA

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