Folia Parasitologica, vol. 59 (2012), issue 4

Folia Parasitologica 59[4] 241-248 (2012) | DOI: 10.14411/fp.2012.033

Molecular survey and microscopic examination of Hepatozoon Miller, 1908 (Apicomplexa: Adeleorina) in lacertid lizards from the western Mediterranean

João P.M.C. Maia1,2, Ana Perera1, D. James Harris1
1 CIBIO-UP, Centro de Investigação em Biodiversidade e Recursos Genéticos da Universidade do Porto, Campus Agrário de Vairão, 4485-661 Vairão, Portugal
2 Departamento de Biologia, Faculdade de Ciências, Universidade do Porto, Rua do Campo Alegre FC4, 4169-007 Porto, Portugal

The genus Hepatozoon Miller, 1908 (Apicomplexa: Adeleorina) is composed of intracellular haemogregarine parasites that are widely distributed among all tetrapod groups. The present study combines microscopic and molecular data on haemogregarine parasites from lizards in the western Mediterranean. We screened tissue samples and examined blood smears for the presence of species of Hepatozoon from four lizards, namely Algyroides marchi Valverde, endemic to Southeast Spain, Podarcis bocagei Seoane from Spain and Portugal, P. hispanica Steindachner from Spain, and P. lilfordi Günther from Cabrera, Balearic Islands (Spain). Our results show that prevalence and intensity of Hepatozoon parasites vary between and within lizard species from different regions. Algyroides marchi and P. bocagei from Spain had the lowest values, whereas P. hispanica had the highest. Phylogeny based on 18S rRNA gene sequences indicates that most of the new Hepatozoon sequences are part of a clade exclusive from North African and Iberian lizards, except for a single P. bocagei isolate that is found related to another clade including isolates from other reptile host species and rodents. Interestingly, isolates from Algyroides form a distinct monophyletic subgroup, which could be a signal of strict host-specificity within this host genus.

Keywords: Apicomplexa, haemogregarine, 18S rRNA, phylogeny, tissue, blood smears

Received: June 7, 2012; Accepted: October 4, 2012; Published: December 1, 2012


References

  1. Abramoff M.D., Magalhaes P.J., Ram S.J. 2004: Image processing with ImageJ. Biophotonics Int. 11: 36-42
  2. Alvarez -Calvo J.A. 1975: Nuevas especies de hemococcidios en lacertidos espanoles. Cuadernos de Cienc. Biol. 4: 207-222
  3. Amo L., Fargallo J.A., Martinez -Padilla J., Millan J., Lopez P., Martin J. 2005b: Prevalence and intensity of blood and intestinal parasites in a field population of a Mediterranean lizard, Lacerta lepida. Parasitol. Res. 96: 413-417 Go to original source... Go to PubMed...
  4. Amo L., Lopez P., Martin J. 2004: Prevalence and intensity of haemogregarinid blood parasites in a population of the Iberian rock lizard, Lacerta monticola. Parasitol. Res. 94: 290-293 Go to original source... Go to PubMed...
  5. Amo L., Lopez P., Martin J. 2005a: Prevalence and intensity of haemogregarine blood parasites and their mite vectors in the common wall lizard, Podarcis muralis. Parasitol. Res. 96: 378-381 Go to original source... Go to PubMed...
  6. Bandelt H.J., Forster P., Roehl A. 1999: Median-joining networks for inferring intraspecific phylogenies. Mol. Biol. Evol. 16: 37-48 Go to original source... Go to PubMed...
  7. Baneth G., Mathew J.S., Shkap V., Macintire D.K., Barta J.R., Ewing S.A. 2003: Canine hepatozoonosis: two disease syndromes caused by separate Hepatozoon spp. Trends Parasitol. 19: 27-31 Go to original source... Go to PubMed...
  8. Barta J.R., Ogedenbre J.D., Martin D.S., Smith T.G. 2012: Phylogenetic position of the adeleorinid coccidia (Myzozoa, Apicomplexa, Coccidia, Eucoccidiorida, Adeleorina) inferred using 18S rDNA sequences. J. Eukaryot. Microbiol. 59: 171-180 Go to original source... Go to PubMed...
  9. Criado -Fornelio A., Buling A., Casado N., Gimenez C., Ruas J., Wendt L., da Rosa-Freitas N., Pinheiro M., Rey-Valeiron C., Barba-Carretero J.C. 2009: Molecular characterization of arthropod-borne hematozoans in wild mammals from Brazil, Venezuela and Spain. Acta Parasitol. 54: 187-193 Go to original source...
  10. Criado -Fornelio A., Martinez -Marcos A., Buling -Sarana A., Barba-Carretero J.C. 2003: Molecular studies on Babesia, Theileria and Hepatozoon in southern Europe: Part II. Phylogenetic analysis and evolutionary history. Vet. Parasitol. 114: 173- 194 Go to original source... Go to PubMed...
  11. Criado -Fornelio A., Rey-Valeiron C., Buling A., Barba-Carretero J.C., Jefferies R., Irwin P. 2007: New advances in molecular epizootiology of canine hematic protozoa from Venezuela, Thailand and Spain. Vet. Parasitol. 144: 261-269 Go to original source... Go to PubMed...
  12. Criado -Fornelio A., Ruas J.L., Casado N., Farias N., Soares P., Muller G., Brum J.G.W., Berne M.E.A., Buling A., BarbaCarretero J.C. 2006: New molecular data on mammalian Hepatozoon species (Apicomplexa: Adeleorina) from Brazil and Spain. J. Parasitol. 92: 93-99 Go to original source... Go to PubMed...
  13. Davis J.R., Boyle S.A., Khan A.A., Gay A.L.J., Grisham J.M., Luque L.E. 2012: Snake parasitism in an urban old-growth forest. Urban Ecosyst. 15: 739-752 Go to original source...
  14. Drummond A.J., Ashton B., Buxton S., Cheung M., Cooper A., Duran C., Field M., Heled J., Kearse M., Markowitz S., Moir R., Stones -Havas S., Sturrock S., Thierer T., Wilson A. 2012: Geneious v5.6. Available from http://www.geneious.com
  15. Felsenstein J. 1985: Confidence and phylogenies: an approach using the bootstrap. Evolution 39: 783-791 Go to original source...
  16. Garcia P., Acedo M.C., Lopez J.J., Sanchis M.C., Morillas F. 1990: Identification of Hepatozoon canis (James, 1905) in Spain. Epidemiological study of an enzootic in La Carolina, Jaen. Investigacion Agraria, Produccion y Sanidad Animales 5: 75-89
  17. Guindon S., Dufayard J.F., Lefort V., Anisimova M., Hordijk W., Gascuel O. 2010: New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Syst. Biol. 59: 307-321 Go to original source... Go to PubMed...
  18. Harris D.J., Maia J.P.M.C., Perera A. 2011: Molecular characterization of Hepatozoon species in reptiles from the Seychelles. J. Parasitol. 97: 106-110 Go to original source... Go to PubMed...
  19. Harris D.J., Maia J.P.M.C., Perera A. 2012: Molecular survey of Apicomplexa in Podarcis wall lizards detects Hepatozoon, Sarcocystis and Eimeria species. J. Parasitol. 98: 592-597 Go to original source... Go to PubMed...
  20. Heneberg P. 2012: On the robustness of phylogenetic analyses: can flatworm 18S rDNA hide between 18S rDNAs of a single mollusc genus? Mol. Phyl. Evol. In Press. DOI: 10.1016/j.ympev.2012.08.003 Go to original source...
  21. Huelsenbeck J.P., Ronquist F. 2001: MrBayes: Bayesian inference of phylogeny. Bioinformatics 17: 754-755 Go to original source... Go to PubMed...
  22. Jovani R., Amo L., Arriero E., Krone O., Marzal A., Shurulinkov P., Tomas G., Sol D., Hagen J., Lopez P., Martin J., Navarro C., Torres J. 2004: Double gametocyte infections in apicomplexan parasites of birds and reptiles. Parasitol. Res. 94: 155-157 Go to original source... Go to PubMed...
  23. Lledo L., Gimenez -Pardo C., Dominguez -Penafiel G., Sousa R., Gegundez M.I., Casado N., Criado A. 2010: Molecular detection of Hemoprotozoa and Rickettsia species in arthropods collected from wild animals in the Burgos province, Spain. VectorBorne Zoon. Dis. 10: 735-738 Go to original source... Go to PubMed...
  24. Maia J.P.M.C., Harris D.J., Perera A. 2011: Molecular survey of Hepatozoon species in lizards from North Africa. J. Parasitol. 97: 513-517 Go to original source... Go to PubMed...
  25. Martin J., Civantos E., Amo L., Lopez P. 2007: Chemical ornaments of male lizards Psammodromus algirus may reveal their parasite load and health state to females. Behav. Ecol. Sociobiol. 62: 173-179 Go to original source...
  26. Mathew J.S., Van Den B ussche R.A., Ewing S.A., Malayer J.R., Latha B.R., Panciera R.J. 2000: Phylogenetic relationships of Hepatozoon (Apicomplexa: Adeleorina) based on molecular, morphologic, and life-cycle characters. J. Parasitol. 86: 366-372 Go to original source... Go to PubMed...
  27. Moco T.C., Silva R.J., Madeira N.G., Paduan K.S., Rubini A.S., Leal D.D.M., O'D wyer L.H. 2012: Morphological, morphometric, and molecular characterization of Hepatozoon spp. (Apicomplexa, Hepatozoidae) from naturally infected Caudisona durissa terrifica (Serpentes, Viperidae). Parasitol. Res. 110: 1393-1401 Go to original source... Go to PubMed...
  28. Morrison D.A. 2009: Evolution of the Apicomplexa: where are we now? Trends Parasitol. 25: 375-382 Go to original source... Go to PubMed...
  29. Ortuno A., Castella J., Criado -Fornelio A., Buling A., BarbaCarretero J.C. 2008: Molecular detection of a Hepatozoon species in stray cats from a feline colony in North-eastern Spain. Vet. J. 177: 134-135 Go to original source... Go to PubMed...
  30. Paterson S., Piertney S.B. 2011: Frontiers in host-parasite ecology and evolution. Mol. Ecol. 20: 869-871 Go to original source... Go to PubMed...
  31. Perkins S.L., Keller A.K. 2001: Phylogeny of nuclear small subunit rRNA genes of hemogregarines amplified with specific primers. J. Parasitol. 87: 870-876 Go to original source... Go to PubMed...
  32. Pinho C., Harris D.J., Ferrand N. 2008: Non-equilibrium estimates of gene flow inferred from nuclear genealogies suggest that Iberian and North African wall lizards (Podarcis spp.) are an assemblage of incipient species. BMC Evol. Biol. 8: 63 Go to original source... Go to PubMed...
  33. Posada D., Crandall K.A. 1998: Modeltest: testing the model of DNA substitution. Bioinformatics 14: 817-818 Go to original source... Go to PubMed...
  34. Poulin R., Mouillot D. 2005: Combining phylogenetic and ecological information into a new index of host specificity. J. Parasitol. 91: 511-514 Go to original source... Go to PubMed...
  35. RD evelopment C ore Team 2009: R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. Available at: http://www.R-project.org
  36. Roca V., Galdon M.A. 2010: Haemogregarine blood parasites in the lizards Podarcis bocagei (Seoane) and P. carbonelli (PerezMellado) (Sauria: Lacertidae) from NW Portugal. Syst. Parasitol. 75: 75-79 Go to original source... Go to PubMed...
  37. Sacchi R., Rubolini D., Gentilli A., Pupin F., Razzetti E., Scali S., Galeotti P., Fasola M. 2007: Morph-specific immunity in male Podarcis muralis. Amphibia-Reptilia 28: 408-412
  38. Sacchi R., Scali S., Cavirani V., Pupin F., Pellitteri -Rosa D., Zuffi M.A.L. 2011: Leukocyte differential counts and morphology from twelve European lizards. Ital. J. Zool. 78: 418-426 Go to original source...
  39. Salkeld D.J., Schwarzkopf L. 2005: Epizootiology of blood parasites in an Australian lizard: a mark-recapture study of a natural population. Int. J. Parasitol. 35: 11-18 Go to original source... Go to PubMed...
  40. Sambrook J., Fritsch E.F., Maniatis T. 1989: Molecular Cloning: A Laboratory Manual. Cold Spring Harbour Press, New York, 545 pp
  41. Santos M.M. de V., O'D wyer L.H., da Silva R.J. 2005: Seasonal variation of Hepatozoon spp. (Apicomplexa, Hepatozoidae) parasitemia from Boa constrictor amarali (Serpentes, Boidae) and Hydrodynastes gigas (Serpentes, Colubridae). Parasitol. Res. 97: 94-97 Go to original source... Go to PubMed...
  42. Smith T.G. 1996: The genus Hepatozoon (Apicomplexa: Adeleina). J. Parasitol. 82: 565-585 Go to original source... Go to PubMed...
  43. Smith T.G., Desser S.S. 1997: Phylogenetic analysis of the genus Hepatozoon Millet 1908 (Apicomplexa: Adeleorina). Syst. Parasitol. 36: 213-221 Go to original source...
  44. Tabar M., Altet L., Francino O., Sanchez A., Ferrer L., Roura X. 2008: Vector-borne infections in cats: molecular study in Barcelona area (Spain). Vet. Parasitol. 151: 332-336 Go to original source... Go to PubMed...
  45. Tamura K., Peterson D., Peterson N., Stecher G., Nei M., Ku mar S. 2011: MEGA5: Molecular Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony methods. Mol. Biol. Evol. 28: 2731-2739 Go to original source... Go to PubMed...
  46. Telford S.R. 2009: Hemoparasites of the Reptilia: Color Atlas and Text. CRC Press, Taylor and Francis Group, Boca Raton, Florida, 394 pp
  47. Telford S.R., Wozniak E.J., Butler J.F. 2001: Haemogregarine specificity in two communities of Florida snakes, with descriptions of six new species of Hepatozoon (Apicomplexa: Hepatozoidae) and possible species of Haemogregarina (Apicomplexa: Haemogregarinidae). J. Parasitol. 87: 890-905 Go to original source... Go to PubMed...
  48. Tome B., Maia J.P.M.C., Harris D.J. 2012: Hepatozoon infection prevalence in four snake genera: influence of diet, prey parasitaemia levels or parasite type? J. Parasitol. 98: 913-917 Go to original source... Go to PubMed...
  49. Ujvari B., Madsen T., Olsson M. 2004: High prevalence of Hepatozoon spp. (Apicomplexa, Hepatozoidae) infection in water pythons (Liasis fuscus) from tropical Australia. J. Parasitol. 90: 670-672 Go to original source... Go to PubMed...
  50. Yabsley M.J., McKibben J., Macpherson C.N., Cattan P.F., Cherry N.A., Hegarty B.C., Breitschwerdt E.B., O'C onnor T., Chandrashekar R., Paterson T., Perea M.L., Ball G., Friesen S., Goedde J., Henderson B., Sylvester W. 2008: Prevalence of Ehrlichia canis, Anaplasma platys, Babesia canis vogeli, Hepatozoon canis, Bartonella vinsonii berkhoffii, and Rickettsia spp. in dogs from Grenada. Vet. Parasitol. 151: 279-285 Go to original source... Go to PubMed...
  51. Zar J.H. 2009: Biostatistical Analysis. Fourth Edition, Prentice Hall, Upper Saddle River, New Jersey, 663 pp